Compounds and methods for inhibiting MRP1

ABSTRACT

The present invention relates to a compound of formula (I), which are useful for inhibiting resistant neoplasms where the resistance is conferred in part or in total by MRP1.

BACKGROUND

[0001] Along with surgery and radiotherapy, chemotherapy continues to bean effective therapy for many cancers. In fact, several types of cancer,such as Hodgkin's disease, large cell lymphoma, acute lymphocyticleukemia, testicular cancer and early stage breast cancer, are nowconsidered to curable by chemotherapy. Other cancers such as ovariancancer, small cell lung and advanced breast cancer, while not yetcurable, are exhibiting positive response to combination chemotherapy.

[0002] One of the most important unsolved problems in cancer treatmentis drug resistance. After selection for resistance to a single cytotoxicdrug, cells may become cross resistant to a whole range of drugs withdifferent structures and cellular targets, e.g., alkylating agents,antimetabolites, hormones, platinum-containing drugs, and naturalproducts. This phenomenon is known as multidrug resistance (MDR). Insome types of cells, this resistance is inherent, while in others, suchas small cell lung cancer, it is usually acquired.

[0003] Such resistance is known to be multifactorial and is conferred byat least two proteins: the 170 kDa P-glycoprotein (MDR1) and the morerecently identified 190 kDa multidrug resistance protein (MRP1).Although both MDR1 and MRP1 belong to the ATP-binding cassettesuperfamily of transport proteins, they are structurally very differentmolecules and share less than 15% amino acid homology. Despite thestructural divergence between the two proteins, by 1994 there were noknown consistent differences in the resistance patterns of MDR1 and MRP1cell lines. However, the association, or lack thereof, of MRP1 andresistance to particular oncolytics is known. See Cole, et. al.,“Pharmacological Characterization of Multidrug Resistant MRP-transfectedHuman Tumor Cells”, Cancer Research, 54:5902-5910, 1994. Doxorubicin,daunorubicin, epirubicin, vincristine, and etoposide are substrates ofMRP1, i.e., MRP1 can bind to these oncolytics and redistribute them awayfrom their site of action, the nucleus, and out of the cell. Id. andMarquardt, D., and Center, M. S., Cancer Research, 52:3157, 1992.

[0004] Doxorubicin, daunorubicin, and epirubicin are members of theanthracycline class of oncolytics. They are isolates of various strainsof Streptomyces and act by inhibiting nucleic acid synthesis. Theseagents are useful in treating neoplasms of the bone, ovaries, bladder,thyroid, and especially the breast. They are also useful in thetreatment of acute lymphoblastic and myeloblastic leukemia, Wilm'stumor, neuroblastoma, soft tissue sarcoma, Hodgkin's andnon-Hodgkin'slymphomas, and bronchogenic carcinoma.

[0005] Vincristine, a member of the vinca alkaloid class of oncolytics,is an isolate of a common flowering herb, the periwinkle plant (Vincarosea Linn). The mechanism of action of vincristine is still underinvestigation but has been related to the inhibition of microtubuleformation in the mitotic spindle. Vincristine is useful in the treatmentof acute leukemia, Hodgkin's disease, non-Hodgkin's malignant lymphomas,rhabdomyosarcoma, neuroblastoma, and Wilm's tumor.

[0006] Etoposide, a member of the epipodophyllotoxin class ofoncolytics, is a semisynthetic derivative of podophyllotoxin. Etoposideacts as a topoisomerase inhibitor and is useful in the therapy ofneoplasms of the testis, and lung.

[0007] Additionally, PCT publications WO99/51236, WO99/51228, andWO99/51227 disclose certain compounds known to be inhibitors of MRP1.

[0008] It is presently unknown what determines whether a cell line willacquire resistance via a MDR1 or MRP1 mechanism. Due to the tissuespecificity of these transporters and/or in the case where one mechanismpredominates or is exclusive, it would be useful to have a selectiveinhibitor of that one over the other. Furthermore, when administering adrug or drugs that are substrates of either protein, it would beparticularly advantageous to coadminister an agent that is a selectiveinhibitor of that protein. It is, therefore, desirable to providecompounds that are selective inhibitors of MDR1 or MRP1.

[0009] This invention relates to novel compounds, which inhibit MRP1 andare therefore useful for the treatment of MRP1 conferred MDR in aresistant neoplasm, or a neoplasm susceptible to resistance in a mammal.

SUMMARY OF THE INVENTION

[0010] The present invention relates to a compound of formula:

[0011] where:

[0012] het is a five (5) membered heteroaryl ring containing N and asecond heteroatom selected from N, O, or S;

[0013] wherein the non-fused carbon atom of the heteroaryl ring isoptionally substituted with R^(b) and; provided that when het ispyrazole or imidazole, the saturated nitrogen of the het ring isoptionally substituted with R^(a);

[0014] wherein R^(b) is C₁-C₆ alkyl, aryl, substituted aryl,heterocycle, substituted heterocycle, an amino acid ester, CH₂OH,CH₂O-heterocycle, halo, CH₂N₃, CH₂SR¹, CH₂NR⁴R⁵, OR¹, SR¹²,S(CH₂)_(n)-phenyl, or NR⁴R⁵, and R^(a) is C₁-C₄ alkyl.

[0015] R is (CH₂)_(m), CHR¹NHR², O(CH₂)₂NHR², (CH₂)_(m), COR³, NHR², and(CH₂)_(m), CHR¹NR⁴R⁵;

[0016] R′ is hydrogen, hydroxy, or O(C₁-C₆ alkyl optionally substitutedwith phenyl or C₃-C₇ cycloalkyl);

[0017] m and m′ are independently at each occurrence 0, 1, or 2;

[0018] R¹ is independently at each occurrence hydrogen or C₁ C₆ alkyl;

[0019] R² is hydrogen, COR⁶, CH₂R^(6′), SO₂R⁷, or a moiety of theformula

[0020] R³ is hydrogen, hydroxy, C₁-C₆ alkoxy, an amino acid ester, anamino acid, or NR⁴R⁵;

[0021] R⁴ is hydrogen or C₁-C₆ alkyl;

[0022] R⁵ is hydrogen, C₁-C₆ alkyl, C₆-C₁₀ bicycloalkyl, (C₁-C₄alkyl)phenyl, (C₁-C₄ alkyl)-CO₂R₁, CH₂CO₂R¹, aryl, substituted aryl,(CH₂)_(n)CHR⁸NHC(O)OC(CH₃)₃, (CH₂)_(n)NH₂, (CH₂)₂NHCOR⁶, (CH)₂OR¹,(CH₂)_(q)-heterocycle, (CH₂)_(q)-substituted heterocycle, or R⁴ and R⁵,together with the nitrogen to which they are attached, combine to form apyrrolidin-1-yl, piperidin-1-yl, hexamethyleneimin-1-yl, ormorpholin-4-yl ring;

[0023] n is 1, 2, 3, or 4;

[0024] q is 0, 1, 2, or 3;

[0025] R⁶ is C₁₋₁₆ alkyl, substituted C₃-C₆ cycloalkyl, aryl,substituted aryl, tert-butoxy, (CH₂)_(q)-heterocycle,(CH₂)_(q)-substituted heterocycle, (CH₂)_(n)S(O)_(r)R¹,C(CH₃)₂CH₂N(R¹)₂, (CH₂)_(n)CHR⁸NHC(O)OC(CH₃)₃, (CH₂)_(n)CHR⁸NH₂,(CH₂)₂NH-aryl, or NHR⁷;

[0026] R^(6′) is C₁-C₆ alkyl, substituted C₃-C₆ cycloalkyl, aryl,substituted aryl, (CH₂)_(q)-heterocycle, (CH₂)_(q)-substitutedheterocycle, (CH₂)_(n)S(O)_(r)R¹, C(CH₃)₂CH₂N(R¹)₂,(CH₂)_(n)CHR⁸NH—C(O)OC(CH₃)₃, (CH₂)_(n)CHR⁸NH₂, or (CH₂)₂NH-aryl;

[0027] r is 0, 1, or 2;

[0028] R⁷ is C₁-C₆ alkyl, phenyl, or substituted phenyl;

[0029] R⁸ is hydrogen or CO₂R¹; and

[0030] R⁹, R¹⁰, and R¹¹ are independently at each occurrence hydrogen,halo, CO₂R¹, aryl, substituted aryl, thiophene, C₁C₄ alkoxy, (C₁C₃alkyl)phenyl, or C₂C₆ alkenyl;

[0031] R¹² is C₁-C₆ alkyl, (C₁-C₄ alkyl)-phenyl, aryl, substituted aryl,heterocycle or substituted heterocycle; or

[0032] a pharmaceutical salt thereof; provided that if R⁹ and R¹⁰ arehydrogen and R¹¹ is chloro, then het is not

[0033] The present invention further relates to a method of inhibitingMRP1 in a mammal which comprises administering to a mammal in needthereof an effective amount of a compound of formula L In anotherembodiment, the present invention relates to a method of inhibiting aresistant neoplasm, or a neoplasm susceptible to resistance in a mammalwhich comprises administering to a mammal in need thereof an effectiveamount of a compound of formula I in combination with an effectiveamount of an oncolytic agent.

[0034] The present invention also relates to a pharmaceuticalformulation comprising a compound of formula I in combination with oneor more oncolytics, pharmaceutical carriers, diluents, or excipientstherefor.

[0035] Furthermore, the invention relates to the use of a compound offormula I, or a pharmaceutically acceptable salt thereof, for themanufacture of a medicament for inhibiting a resistant neoplasm, or aneoplasm susceptible to resistance in a mammal.

[0036] Furthermore, the invention relates to the use of a compound offormula I, or a pharmaceutically acceptable salt thereof, for themanufacture of a medicament for inhibiting MRP1.

[0037] Furthermore, the invention relates to the use of a compound offormula I, or a pharmaceutically acceptable salt thereof, for themanufacture of a medicament for inhibiting MRP1 conferred MDR in aresistant neoplasm, or a neoplasm susceptible to resistance in a mammal.

[0038] Furthermore, the invention relates to the use of a compound offormula I in therapy.

DETAILED DESCRIPTION OF THE INVENTION

[0039] The current invention concerns the discovery that compounds offormula I are selective inhibitors of multidrug resistant protein(MRP1), and are thus useful in treating MRP1 conferred multidrugresistance (MDR) in a resistant neoplasm and a neoplasm susceptible toresistance.

[0040] The terms “inhibit” as it relates to MRP1 and “inhibiting MRP1”refer to prohibiting, alleviating, ameliorating, halting, restraining,slowing or reversing the progression of, or reducing MRP1's ability toredistribute an oncolytic away from the oncolytic's site of action, mostoften the neoplasm's nucleus, and out of the cell.

[0041] As used herein, the term “effective amount of a compound offormula I” refers to an amount of a compound of the present inventionwhich is capable of inhibiting MRP1. The term “effective amount of anoncolytic agent” refers to an amount of oncolytic agent capable ofinhibiting a neoplasm, resistant or otherwise.

[0042] The term “inhibiting a resistant neoplasm, or a neoplasmsusceptible to resistance” refers to prohibiting, halting, restraining,slowing or reversing the progression of, reducing the growth of, orkilling resistant neoplasms and/or neoplasms susceptible to resistance.

[0043] The term “resistant neoplasm” refers to a neoplasm, which isresistant to chemotherapy where that resistance is conferred in part, orin total, by MRP1. Such neoplasms include, but are not limited to,neoplasms of the bladder, bone, breast, lung (small-cell), testis, andthyroid and also includes more particular types of cancer such as, butnot limited to, acute lymphoblastic and myeloblastic leukemia, Wilm'stumor, neuroblastoma, soft tissue sarcoma, Hodgkin's and non-Hodgkin'slymphomas, and bronchogenic carcinoma.

[0044] A neoplasm, which is “susceptible to resistance”, is a neoplasmwhere resistance is not inherent nor currently present but can beconferred by MRP1 after chemotherapy begins. Thus, the methods of thisinvention encompass a prophylactic and therapeutic administration of acompound of formula I.

[0045] The term “chemotherapy” refers to the use of one or moreoncolytic agents where at least one oncolytic agent is a substrate ofMRP1. A “substrate of MRP1” is an oncolytic that binds to MRP1 and isredistributed away from the oncolytics site of action (the nucleus ofthe neoplasm) and out of the cell, thus, rendering the therapy lesseffective. Preferred oncolytic agents are doxorubicin, daunorubicin,epirubicin, vincristine, and etoposide.

[0046] The terms “treat” or “treating” bear their usual meaning whichincludes preventing, prohibiting, alleviating, ameliorating, halting,restraining, slowing or reversing the progression, or reducing theseverity of MRP1 derived drug resistance in a multidrug resistant tumor.

[0047] In the general formulae of the present document, the generalchemical terms have their usual meanings. For example, the term “C₁-C₄alkyl” refers to methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl,cyclobutyl, s-butyl, and t-butyl. The term “C₁-C₆ alkyl” refers to amonovalent, straight or branched saturated hydrocarbon containing from 1to 6 carbon atoms. Additionally, the term “C₁-C₆ alkcyl” includes C₁-C₄alkyl groups and C₃-C₆ cycloalkyls. The term “C₁-C₆ alkyl” includes, butis not limited to, cyclopentyl, pentyl, hexyl, cyclohexyl, and the like.The term “C₃-C₆ cycloalkyl” refers to cyclopropyl, cyclobutyl,cyclopentyl, and cyclohexyl. The term “C₅-C₇ cycloalkyl” refers tocyclopentyl, cyclohexyl, and cycloheptyl. The term “C₆C₁₀ bicycloalkyl”refers to bicyclo-[2.1.1]hexanyl, [2.2.1]heptanyl, [3.2.1]octanyl,[2.2.2]octanyl, [3.2.2]nonanyl, [3.3.1]nonanyl, [3.3.2]decanyl, and[4.3.1]decanyl ring systems.

[0048] The terms “C₁-C₄ alkoxy” and “C₁-C₆ alkoxy” refer to moieties ofthe formula O—(C₁-C₄ alkyl) and O—(C₁-C₆ alkyl) respectively.

[0049] The term “substituted C₃-C₆ cycloalkyl” refers to a C₃-C₆cycloalkyl substituted once with a phenyl, substituted phenyl, or CO₂R¹group.

[0050] The term “halo” or “halide” refers to fluoro, chloro, bromo, andiodo.

[0051] The term “aryl” refers to phenyl, benzyl, and naphthyl.

[0052] The terms “substituted aryl” refers to a phenyl, benzyl, andnaphthyl group respectively substituted from 1 to 3 times independentlywith C₁-C₆ alkyl, C₁-C₄ alkoxy, halo, hydroxy, trifluoromethyl, N(R¹)₂,SO₂N(R¹)₂, NH-Pg, C₁-C₆ alkoxy, benzyloxy, CO₂R¹, C₅-C₇ cycloalkyl,trifluoromethoxy, or nitro.

[0053] The term “heterocycle” is taken to mean stable unsaturated andsaturated 5- and 6-membered rings containing from 1 to 3 heteroatomsselected from the group consisting of nitrogen, oxygen and sulfur, saidrings being optionally benzofused. All of these rings may be substitutedwith up to three substituents independently selected from the groupconsisting of halo, C₁-C₄ alkoxy, C₁-C₄ alkyl, cyano, nitro, hydroxy,—S(O)_(m)—(C₁-C₄ alkyl) and —S(O)_(m)-phenyl where m is 0, 1 or 2.Saturated rings include, for example, pyrrolidinyl, piperidinyl,piperazinyl, tetrahydrofuryl, oxazolidinyl, dioxanyl, pyranyl, and thelike. Benzofused saturated rings include indolinyl,1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl and thelike. Unsaturated rings include furyl, thienyl, pyridinyl, pyrrolyl,N-methylpyrrolyl, oxazolyl, isoxazolyl, pyrazolyl, imidazolyl,triazolyl, oxadiazolyl, thiadiazolyl, thiazolyl, pyrimidinyl, pyrazinyl,pyridazinyl, and the like. Benzofused unsaturated rings includeisoquinolinyl, benzoxazolyl, benzthiazolyl, quinolinyl, benzofuranyl,thionaphthyl, indolyl and the like.

[0054] The term “heteroaryl” is taken to mean an unsaturated orbenzofused unsaturated heterocycle as defined in the previous paragraph.

[0055] The term “substituted heterocycle” refers to a heterocyclic ringsubstituted 1 or 2 times independently with a C₁-C₆ alkyl, halo, benzyl,phenyl, trifluoromethyl. Saturated heterocyclic rings may beadditionally substituted 1 or 2 times with an oxo group, however, totalsubstitution of the saturated heterocyclic ring may not exceed twosubstituents.

[0056] The term “amino acid” refers to a chemical unit made up of both abasic amino group and an acidic carboxyl group. Examples of amino acidsinclude alanine, asparagine, cysteine, glutamine, glycine, isoleucine,leusine, methionine, phenylalanine, proline, serine threonine,tryptophan, tyrosine, valine, aspartic acid, glutamic acid, arginine,histidine, and lysine.

[0057] The term “amino acid ester” as used in this specification refersto an amino acid where the carboxy group is substituted with a C₁-C₆alkyl group. That is, the alkyl group when taken together with thecarboxy group forms a C₁-C₆ alkyl ester. A skilled artisan wouldappreciate that some amino acids have two carboxy groups that may besubstituted with a C₁-C₆ alkyl group, for example, aspartic acid andglutamic acid. This invention contemplates the possibility of amino acidmono- or diesters in these circumstances.

[0058] The term “protecting group” (Pg) refers to an amino protectinggroup or a hydroxy protecting group. The species of protecting groupwill be evident from whether the “Pg” group is attached to a nitrogenatom (amino protecting group) or attached to an oxygen atom (hydroxyprotecting group).

[0059] The term “amino protecting group” as used in this specificationrefers to a substituent(s) of the amino group commonly employed to blockor protect the amino functionality while reacting other functionalgroups on the compound. Examples of such amino-protecting groups includethe formyl group, the trityl group, the phthalimido group, the acetylgroup, the trichloroacetyl group, the chloroacetyl, bromoacetyl, andiodoacetyl groups, urethane-type blocking groups such asbenzyloxycarbonyl, 9-fluorenylmethoxycarbonyl (“FMOC”), and the like;and like amino protecting groups. The species of amino protecting groupemployed is not critical so long as the derivitized amino group isstable to the condition of subsequent reaction(s) on other positions ofthe molecule and can be removed at the appropriate point withoutdisrupting the remainder of the molecule. Similar amino protectinggroups used in the cephalosporin, penicillin, and peptide arts are alsoembraced by the above terms. Further examples of groups referred to bythe above terms are described by T. W. Greene, “Protective Groups inOrganic Synthesis”, John Wiley and Sons, New York, N.Y., 1991, Chapter 7hereafter referred to as “Greene”. A preferred amino protecting group ist-butyloxycarbonyl.

[0060] The term “hydroxy protecting group” denotes a group understood byone skilled in the organic chemical arts of the type described inChapter 2 of Greene. Representative hydroxy protecting groups include,for example, ether groups including methyl and substituted methyl ethergroups such as methyl ether, methoxymethyl ether, methylthiomethylether, tert-butylthiomethyl ether, (phenyldimethylsilyl)methoxy-methylether, benzyloxymethyl ether, p-methoxybenzyloxy-methyl ether, andtert-butoxymethyl ether, substituted ethyl ether groups such asethoxyethyl ether, 1-(2-chloroethoxy)ethyl ether,2,2,2-trichloroethoxymethyl ether, and 2-(trimethylsilyl)ethyl ether,isopropyl ether groups; phenyl and substituted phenyl ether groups suchas phenyl ether, p-chlorophenyl ether, p-methoxyphenyl ether, and2,4-dinitrophenyl ether, benzyl and substituted benzyl ether groups suchas benzyl ether, p-methoxybenzyl ether, o-nitrobenzyl ether, and2,6-dichlorobenzyl ether; and alkylsilyl ether groups such astrimethyl-triethyl- and triisopropylsilyl ethers, mixed alkylsilyl ethergroups such as dimethylisopropylsilyl ether, and diethylisopropylsilylether, and ester protecting groups such as formate ester, benzylformateester, mono-, di-, and trichloroacetate esters, phenoxyacetate ester,and p-chlorophenoxyacetate and the like. The species of hydroxyprotecting group employed is not critical so long as the derivatizedhydroxy group is stable to the conditions of subsequent reaction(s) onother positions of the intermediate molecule and can be selectivelyremoved at the appropriate point without disrupting the remainder of themolecule including any other hydroxy protecting group(s).

[0061] The term “carbonyl activating group” refers to a substituent of acarbonyl that increases the susceptibility of that carbonyl tonucleophilic addition. Such groups include, but are not limited to,alkoxy, aryloxy, nitrogen containing unsaturated heterocycles, or aminogroups such as oxybenzotriazole, imidazolyl, nitrophenoxy,pentachloro-phenoxy, N-oxysuccinimide, N,N′-dicyclohexylisoure-O-yl,N-hydroxy-N-methoxyamino, and the like; acetates, formates, sulfonatessuch as methanesulfonate, ethanesulfonate, benzenesulfonate, orp-toluenylsulfonate, and the like; and halides especially chloride,bromide, or iodide.

[0062] The term “carbonyl activating reagent” refers to a reagent thatconverts the carbonyl of a carboxylic acid group to one that is moreprone to nucleophilic addition and includes, but is not limited to, suchreagents as those found in “The Peptides”, Gross and Meienhofer, Eds.,Academic Press (1979), Ch. 2 and M. Bodanszky, “Principles of PeptideSynthesis”, 2^(nd) Ed., Springer-Verlag Berlin Heidelberg, 1993,hereafter referred to as “The Peptides” and “Peptide Synthesis”respectively. Specifically, carbonyl activating reagents include thionylbromide, thionyl chloride, oxalyl chloride, and the like; alcohols suchas nitrophenol, pentachlorophenol, and the like; amines such asN-hydroxy-N-methoxyamine and the like; acid halides such as acetic,formic, methanesulfonic, ethanesulfonic, benzenesulfonic, orp-tolylsulfonic acid halide, and the like; and compounds such as1,1′-carbonyldiimidazole, benzotriazole, imidazole,N-hydroxysuccinimide, dicyclohexylcarbodiimide, and the like.

[0063] In general, the term “pharmaceutical” when used as an adjectivemeans substantially non-toxic to living organisms. For example, the term“pharmaceutical salt” as used herein, refers to salts of the compoundsof formula I which are substantially non-toxic to living organisms. See,e.g., Berge, S. M, Bighley, L. D., and Monkhouse, D. C., “PharmaceuticalSalts”, J. Pharm. Sci., 66:1, 1977. Typical pharmaceutical salts includethose salts prepared by reaction of the compounds of formula I with aninorganic or organic acid or base. Such salts are known as acid additionor base addition salts respectively. These pharmaceutical saltsfrequently have enhanced solubility characteristics compared to thecompound from which they are derived, and thus are often more amenableto formulation as liquids or emulsions.

[0064] The term “acid addition salt” refers to a salt of a compound offormula I prepared by reaction of a compound of formula I with a mineralor organic acid. For exemplification of pharmaceutical acid additionsalts see, e.g., Berge, S. M, Bighley, L. D., and Monkhouse, D. C., J.Pharm Sci., 66:1, 1977. Since compounds of this invention can be basicin nature, they accordingly react with any of a number of inorganic andorganic acids to form pharmaceutical acid addition salts.

[0065] The pharmaceutical acid addition salts of the invention aretypically formed by reacting the compound of formula I with an equimolaror excess amount of acid. The reactants are generally combined in amutual solvent such as diethylether, tetrahydrofuran, methanol, ethanol,isopropanol, benzene, and the like. The salts normally precipitate outof solution within about one hour to about ten days and can be isolatedby filtration or other conventional methods.

[0066] Acids commonly employed to form acid addition salts are inorganicacids such as hydrochloric acid, hydrobromic acid, hydroiodic acid,sulfuric acid, phosphoric acid, and the like, and acids commonlyemployed to form such salts are inorganic acids such as hydrochloricacid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid,and the like, and organic acids, such as p-toluenesulfonic acid,methanesulfonic acid, oxalic acid, p-bromophenylsulfonic acid, carbonicacid, succinic acid, citric acid, benzoic acid, acetic acid and thelike. Examples of such pharmaceutically acceptable salts thus are thesulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate,monohydrogenphosphate, dihydrogenphosphate, metaphosphate,pyrophosphate, chloride, bromide, iodide, acetate, propionate,decanoate, caprylate, acrylate, formate, isobutyrate, caproate,heptanoate, propiolate, oxalate, malonate, succinate, suberate,sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate,benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate,hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, xylenesulfonate,phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate,β-hydroxybutyrate, glycollate, tartrate, methanesulfonate,propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate,mandelate and the like.

[0067] The term “base addition salt” refers to a salt of a compound offormula I prepared by reaction of a compound of formula I with a mineralor organic base. For exemplification of pharmaceutical base additionsalts see, e.g., Berge, S. M, Bighley, L. D., and Monkhouse, D. C., J.Pharm. Sci., 66:1, 1977. This invention also contemplates pharmaceuticalbase addition salts of compounds of formula I. The skilled artisan wouldappreciate that some compounds of formula I may be acidic in nature andaccordingly react with any of a number of inorganic and organic bases toform pharmaceutical base addition salts. Examples of pharmaceutical baseaddition salts are the ammonium, lithium, potassium, sodium, calcium,magnesium, methylamino, diethylamino, ethylene diamino, cyclohexylamino,and ethanolamino salts, and the like of a compound of formula I.

[0068] While all of the compounds of the present invention are useful,certain of the compounds are particularly interesting and are preferred.The following listing sets out several groups of preferred compounds. Itwill be understood that each of the listings may be combined with otherlistings to create additional groups of preferred embodiments.

[0069] i. m is 0;

[0070] ii. R is at the meta position;

[0071] iii. R is (CH₂)_(m′)CHR¹NHR²;

[0072] iv. R is (CH₂)_(m′)COR³;

[0073] v. R is (CH₂)_(m′)CHR¹NR⁴R⁵;

[0074] vi. m′ is 0;

[0075] vii. m′ is 1;

[0076] viii. R¹ is methyl;

[0077] ix. R¹ is hydrogen;

[0078] x. R² is 3,4,5-trimethoxyphenylmethyl;

[0079] xi. R³ is (3,4,5-trimethoxyphenyl)amino;

[0080] xii. R³ is (4-aminosulfonylphenyl)amino;

[0081] xiii. R³ is (6-methoxyquinolin-8-yl)amino;

[0082] xiv. R⁴ is hydrogen;

[0083] xv. R⁵ is 5-methylisoxazol-3-yl;

[0084] xvi. R⁵ is 3,5-dimethoxy-4-hydroxybenzyl;

[0085] xvii. R⁵ is 3,4,5-trimethoxyphenyl;

[0086] xviii. R′ is hydrogen;

[0087] xix. R⁹ is hydrogen;

[0088] xx. R⁹ is halo;

[0089] xxi. R¹⁰ is hydrogen;

[0090] xxii. R¹¹ is halo;

[0091] xxiii. R¹¹ is C₁-C₄ alkoxy;

[0092] xxiv. R¹¹ is optionally substituted aryl;

[0093] xxv. R is (CH₂)_(m′)CHR¹NHR²;

[0094] xxvi. R is O(CH₂)₂NHR²

[0095] xxvii. R is (CH₂)_(m′)COR³;

[0096] xxviii. R is NHR²;

[0097] xxix. R is (CH₂)_(m′)CHR¹NR⁴R⁵;

[0098] xxx. R′ is hydroxy;

[0099] xxxi. R′ is O(C₁-C₆ alkyl optionally substituted with phenyl orC₃C₇ cycloalkyl);

[0100] xxxii. m is 1;

[0101] xxxiii. m is 2;

[0102] xxxiv. m′ is 2;

[0103] xxxv. R¹ is C₁-C₆ alkyl;

[0104] xxxvi. R² is hydrogen;

[0105] xxxvii. R² is COR⁶;

[0106] xxxviii. R² is CH₂R^(6′);

[0107] xxxix. R² is SO₂R⁷;

[0108] xl. R² is a moiety of the formula

[0109] xli. R³ is hydrogen;

[0110] xlii. R³ is hydroxy;

[0111] xliii. R³ is C₁-C₆ alkoxy;

[0112] xliv. R³ is an amino acid ester,

[0113] xlv. R³ is an amino acid;

[0114] xlvi. R³ is NR⁴R⁵;

[0115] xlvii. R⁴ is C₁-C₆ alkyl;

[0116] xlviii. R⁵ is hydrogen;

[0117] xlix. R⁵ is C₁-C₆ alkyl;

[0118] l. R⁵ is C₆-C₁₀ bicycloalkyl;

[0119] li. R⁵ is (C₁-C₄ alkyl)phenyl;

[0120] lii. R⁵ is (C₁-C₄ alkyl)-CO₂R¹;

[0121] liii. R⁵ is CH₂CO₂R¹;

[0122] liv. R⁵ is aryl;

[0123] lv. R⁵ is substituted aryl;

[0124] lvi. R⁵ is (CH₂)_(n)CHR⁸NHC(O)OC(CH₃)₃;

[0125] lvii. R⁵ is (CH₂)_(n)NH₂;

[0126] lviii. R⁵ is (CH₂)₂NHCOR⁶;

[0127] lix. R⁵ is (CH₂)₂OR¹;

[0128] lx. R⁵ is (CH₂)_(q)-heterocycle;

[0129] lxi. R⁵ is (CH₂)_(q)-substituted heterocycle;

[0130] lxii. R⁴ and R⁵, together with the nitrogen to which they areattached, combine to form a pyrrolidin-1-yl, piperidin-1-yl,hexamethyleneimin-1-yl, or morpholin-4-yl ring;

[0131] lxiii. n is 1;

[0132] lxiv. n is 2;

[0133] lxv. n is 3;

[0134] lxvi. n is 4;

[0135] lxvii. q is 0;

[0136] lxviii. q is 1;

[0137] lxix. q is 2;

[0138] lxx. q is 3;

[0139] lxxi. R⁶ is C₁C₆ alkyl;

[0140] lxxii. R⁶ is substituted C₃-C₆ cycloalkyl;

[0141] lxxiii. R⁶ is aryl;

[0142] lxxiv. R⁶ is substituted aryl;

[0143] lxxv. R⁶ is tert-butoxy;

[0144] lxxvi. R⁶ is (CH₂)_(q)-heterocycle;

[0145] lxxvii. R⁶ is (CH₂)_(q)-substituted heterocycle;

[0146] lxxviii. R⁶ is (CH₂)S(O₅R¹;

[0147] lxxix. R⁶ is C(CH₃)₂CH₂N(R)₂;

[0148] lxxx. R⁶ is (CH₂S)CHR⁸NHC(O)OC(CH₃)₃;

[0149] lxxxi. R⁶ is (CH₂)_(n)CHR⁸NH₂;

[0150] lxxxii. R⁶ is (CH₂)₂NH-aryl;

[0151] lxxxiii. R⁶ is NHR⁷;

[0152] lxxxiv. R^(6′) is C₁-C₆ alkyl;

[0153] lxxxv. R^(6′) is substituted C₃-C₆ cycloalkyl;

[0154] lxxxvi. R^(6′) is aryl;

[0155] lxxxvii. R^(6′) is substituted aryl;

[0156] lxxxviii. R^(6′) is (CH₂)_(q)-heterocycle;

[0157] lxxxix. R^(6′) is (CH₂)_(q)-substituted heterocycle;

[0158] xc. R^(6′) is (CH₂)_(n)S(O)_(r)R¹;

[0159] xci. R^(6′) is C(CH₃₂CH₂N(R¹)₂;

[0160] xcii. R^(6′) is (CH₂)_(n)CHR⁸NHC(O)OC(CH₃)₃;

[0161] xciii. R^(6′) is (CH₂)_(n)CHR⁸NH₂;

[0162] xciv. R^(6′) is (CH₂)₂NH-aryl;

[0163] xcv. r is 0;

[0164] xcvi. r is 1;

[0165] xcvii. r is 2;

[0166] xcviii. R⁷ is C₁-C₆ alkyl;

[0167] xcix. R⁷ is phenyl;

[0168] c. R⁷ is substituted phenyl;

[0169] ci. R⁸ is hydrogen;

[0170] cii. R⁸ is CO₂R¹;

[0171] ciii. R⁹ is CO₂R¹;

[0172] civ. R⁹ is aryl;

[0173] cv. R⁹ is substituted aryl;

[0174] cvi. R⁹ is thiophene;

[0175] cvii. R⁹ is C₁-C₄ alkoxy;

[0176] cviii. R⁹ is (C₁-C₃ alkyl)-phenyl;

[0177] cix. R⁹ is C₂-C₆ alkenyl;

[0178] cx. R¹⁰ is halo;

[0179] cxi. R¹⁰ is CO₂R¹;

[0180] cxii. R¹⁰ is aryl;

[0181] cxiii. R¹⁰ is substituted aryl;

[0182] cxiv. R¹⁰ is, thiophene;

[0183] cxv. R¹⁰ is C₁-C₄ alkoxy;

[0184] cxvi. R¹⁰ is (C₁-C₃ alkyl)-phenyl;

[0185] cxvii. R¹⁰ is C₂-C₆ alkenyl;

[0186] cxviii. R¹¹ is hydrogen;

[0187] cxix. R¹ is CO₂R¹;

[0188] cxx. R¹ is aryl;

[0189] cxxi. R¹¹ is substituted aryl;

[0190] cxxii. R¹¹ is thiophene;

[0191] cxxiii. R¹¹ is C₁-C₄ alkoxy;

[0192] cxxiv. R¹¹ is (C₁-C₃ alkyl)phenyl;

[0193] cxxv. R¹¹ is C₂-C₆ alkenyl;

[0194] cxxvi. R¹² is C₁-C₆ alkyl;

[0195] cxxvii. R¹² is (C₁-C₄ alkyl)phenyl;

[0196] cxxviii. R¹² is aryl;

[0197] cxxix. R¹² is substituted aryl;

[0198] cxxx. R¹² is heterocycle;

[0199] cxxxi. R¹² is substituted heterocycle;

[0200] cxxxii. Het is

[0201] cxxxiii. Het is

[0202] cxxxiv. Het is

[0203] cxxxv. Het is

[0204] cxxxvi. Het is

[0205] cxxxvii. Het is

[0206] cxxxviii. Het is

[0207] cxxxix. R^(b) is C₁-C₆ alkyl;

[0208] cxl. R^(b) is aryl;

[0209] cxli. R^(b) is halo;

[0210] cxlii. R^(b) is SR¹²;

[0211] cxliii. R^(b) is optionally substituted heterocycle;

[0212] cxliv. R^(b) is —NR⁴R⁵;

[0213] cxlv. R^(a) is t-butyl;

[0214] cxlvi. The compound is a pharmaceutical salt; and

[0215] cxlvii. The compound is the hydrochloride salt.

[0216] The compounds of the present invention can be prepared by avariety of procedures, some of which are illustrated in the Schemesbelow. The particular order of steps required to produce the compoundsof formula I is dependent upon the particular compound beingsynthesized, the starting compound, and the relative lability of thesubstituted moieties.

[0217] Compounds of formula I(a), wherein R^(b), the substituent fromthe non-fused carbon, is C₁-C₆ alkyl, optionally substituted aryl,CH₂OH, or CH₂O-heterocycle may be prepared from compounds of formulaII(a) as illustrated in Scheme 1 below where R, R′, R⁹, R¹⁰, R¹¹, hetand m are as described supra.

[0218] Compounds of formula I(a) may be prepared by dissolving orsuspending a compound of formula II(a) in a suitable solvent, preferablydimethylformamide, and adding a suitable base, including potassiummethoxide, potassium tert-butoxide and preferably potassium carbonate.The base is typically employed in a large molar excess, usually in abouta 4 to about an 8 molar excess relative to the compound of formulaII(a). Preferably, about a 5 to about a 7 molar excess is employed.

[0219] The reactants are typically combined at a temperature from about0° C. to about 100° C. When het is isoxazole, oxazole, or imidazole, thereactants are preferably combined at room temperature and the resultingsolution is typically heated from about 30° C. to about the refluxtemperature of the mixture for from 30 minutes to about 18 hours.Preferably, the mixture is heated to at least 50° C. for from about 1 toabout 6 hours, and is most preferably heated to from about 65° C. toabout 75° C. for from about 1.5 hours to about 3 hours. When het ispyrazole the reactants are preferably combined at room temperature andthe resulting solution is typically heated to about 100° C. for from 30minutes to about 18 hours.

[0220] Any protecting groups remaining in the cyclized compound offormula I may be removed as taught in Greene to provide the compounds offormula I. Preferred choices of protecting groups and methods for theirremoval may be found in the Preparations and Examples sections below.

[0221] Compounds of formula I(b), wherein het is substituted with halomay be prepared from compounds of formula II(b) as illustrated in Scheme2 below where R, R′, R⁹, R¹⁰, R¹¹, het and m are as described supra.

[0222] Compounds of formula I(b) may be prepared by dissolving orsuspending a compound of formula II(b) in a suitable solvent and addinga suitable base, in an inert atmosphere, preferably under N₂. Typicallya preferred and convenient solvent is dimethylformamide. A preferredbase is sodium trimethylsilanolate. The base is typically employed in aslight molar excess, usually in about a 1.05 molar excess relative tothe compound of formula I(b). The reactants are typically combineddropwise at room temperature over a period of time from about 2 hours toabout 4 hours.

[0223] The skilled artisan would appreciate that if other bases are usedin the reaction of Scheme 2, the substituent of the het functionalitymay change. For example if sodium methylthiolate is used as thepreferred base, the compound of formula I(b) will be converted into thecompound of formula I wherein the het functionality is substituted with—SCH₃.

[0224] Additionally, the compound of formula I(b) can be preparedaccording to Scheme 1 wherein the reactants are combined at 0° C. andmixed at −10° C. for approximately three hours. The solution is thenwarmed to room temperature and mixed for an additional 2 to 3 hours.

[0225] Certain compounds of formula I(a) are useful MRP1 inhibitors andare also useful intermediates for the preparation of other compounds offormula I. As is shown in Scheme 3, when R^(b) of formula I(a) isCH₂O-Pg, wherein “Pg” is a protecting group, the compound of formulaI(a) may be further reacted by methods known in the art to producecompounds of formula I(c) where R^(b) is CH₂OH, CH₂N₃, CH₂SR¹, orCH₂NR⁴R⁵, and R, R′, R¹, R⁴, R⁵, R⁹, R¹⁰, R¹¹, het, and m are as definedabove.

[0226] Compounds of formula I(c) wherein het is substituted with CH₂OHmay be prepared by dissolving or suspending a compound of formula I(a)in a suitable solvent and adding a suitable acid. Typically a preferredand convenient solvent is methanol/dichloromethane (2:1). A preferredacid is p-toluenesulfonic acid hydrate. The acid is typically employedin a slight molar excess, usually in about a 1.05 molar excess relativeto the compound of formula I(a). The reactants are typically combined atroom temperature and mixed from about 1 hour to about 3 hours.

[0227] The skilled artisan would appreciate that the alcohol can befurther converted to compounds of formula I(c) where het is substitutedwith CH₂N₃, CH₂SR¹, or CH₂NR⁴R⁵ by methods well known in the art. Forgeneral examples of these procedures, see the Preparations and Examplesection.

[0228] Compounds of formula I(b) when het is substituted with chloro areuseful MRP1 inhibitors and are also useful intermediates for thepreparation of other compounds of formula I. As is shown in Scheme 4,the compound of formula I(b) may be further reacted with a nucleophileby methods known in the art to produce compounds of formula I(d) wherehet is substituted with an amino acid ester, OR¹, SR¹²,S(CH₂)_(n)-phenyl, NR⁴R⁵, or an optionally substituted heterocycleattached via a heteroatom and R, R′, R⁹, R¹⁰, R¹¹, R¹, R¹², n, het and mare as defined above.

[0229] Compounds of formula I(o) may be prepared by dissolving orsuspending a compound of formula I(b) in a suitable solvent and addingan appropriate nucleophile, in an inert atmosphere, preferably under N₂.Typically a preferred and convenient solvent is dimethylformamide. Thenucleophile is typically employed in a molar excess, usually in about a2 to about a 4 molar excess relative to the compound of formula I(b).

[0230] The reactants are preferably combined at room temperature and theresulting solution is typically mixed for from about 30 minutes to about3 hours, until the reaction is complete as measure by the consumption ofthe compound of formula I(b). The skilled artisan would appreciate thatthe reaction, depending on the nucleophile used, may require more timeto react and may, also require heating. In these instances, it ispreferred to mix the reactants for from approximately 15 toapproximately 20 hours, then heat the solution to from about 50° C. toabout 80° C. and mix for an additional 3 hours or until the reaction iscomplete as measure by the consumption of the compound of formula I(b).

[0231] Compounds of formula I where R is (CH₂)_(m′)CHR¹NR⁴R⁵ may beprepared from compounds of formula I(n) as illustrated in Scheme 5 belowwhere m, m′, R′, R^(a), R^(b), R¹, R⁴, R⁵, R⁹, R¹⁰, and 1 are asdescribed supra.

[0232] The compounds of formula I(n) may be reductively aminated to formthe compounds of formula I(e). Reductive aminations are well knowntransformations, see, e.g., Larock, “Comprehensive OrganicTransformations”, pg. 421, VCH Publishers, New York, N.Y., 1989,hereafter referred to as “sock”.

[0233] Amines of formula m may be dissolved or suspended in a suitablesolvent, optionally in the presence of a suitable base, preferablyN-methyl morpholine or triethylamine, when the compound of formula m isan acid addition salt to convert the salt to its free amine form, and acompound of formula I(n) is added. A Lewis acid catalyst, such astitanium(IV) isopropoxide, may optionally be employed. Once it isdetermined that the compound of formula I(d) has been substantiallyconsumed, the intermediate is typically reacted in situ with a suitablereducing agent to provide the compounds of formula I(e). The overallconversion may be performed at about 0° C. to the boiling point of themixture, but room temperature is a preferred reaction temperature. Theformation of the compounds of formula I(e) may take from 15 minutes to24 hours as measure by the consumption of the compound of formula I(d).Methanol is typically a preferred solvent.

[0234] Suitable reducing agents include, but are not limited to,hydrogen over palladium or platinum on carbon, borane or complexes ofborane, e.g., borane-pyridine, borane-butylamine, andborane-dimethylamine complex; and borohydride reducing agents such assodium borohydride or sodium cyanoborohydride. Sodium cyanoborohydrideis a preferred reducing agent.

[0235] Compounds of formula I where R is (CH₂)_(m′)CHR¹NHR² orO(CH₂)₂NHR² and R² is CH₂R^(6′) may be prepared from compounds offormula 1(f) and 1(h) as illustrated in Scheme 6 below where X is halideand m, m′, R′, R^(a), R^(b), R¹, R^(6′), R⁹, R¹⁰, and R¹¹ are asdescribed supra.

[0236] The compounds of formulas I(f) and I(h) may be reductivelyalkylated to form the corresponding compounds of formulas I(g) and I(i),respectively. Reductive alkylation of primary amines are well knowntransformations, see, e.g., Larock, pg. 434-435

[0237] The skilled artisan will appreciate that the treatment of acylhalides with amines is a very general reaction for the preparation ofamides, see, e.g. March J, Advanced Organic Chemistry, 1985, 3rdedition, page 370. The reaction is highly exothermic and must becarefully controlled, usually by cooling.

[0238] Once it is determined that the compound of formula IV has beensubstantially consumed, the intermediate is typically reacted in situwith a suitable reducing agent to provide the compounds of formula I(g)and I(i), respectively. The overall conversion may be performed at about0° C. to the boiling point of the mixture but room temperature is apreferred reaction temperature. The formation of the compounds offormulas I(g) and I(i) may take from 15 minutes to 24 hours as measureby the consumption of the compound of formula IV.

[0239] A base is typically employed when the compound of formula I(f) or1(h) is an acid addition salt in order to convert the salt to its freeamine form. Preferred bases for this purpose are N-methylmorpholine andtriethylamine. A preferred Lewis acid catalyst is titanium(V)isopropoxide. Suitable reducing agents include, but are not limited to,borane or complexes of borane, e.g., borane-pyridine,borane-t-butylamine, and borane-dimethylamine complex; and lithiumaluminum hydride.

[0240] The skilled artisan would appreciate that the compounds offormulas I(f) and I(h) may be converted to the corresponding compoundsof formulas I(g) and I(i), respectively, by reacting with ketones andaldehydes as is represented in Scheme 5. Reductive alkylation of primaryamines are well known transformations, see, e.g., Larock, supra.

[0241] Compounds of formula 1(h) may be converted to other compounds ofthe invention by methods well known in the chemical arts. Compounds offormula I where R is O(CH₂)₂NHR², and R² is CONHR⁷ or a moiety of theformula

[0242] and m, R′, R⁷, R⁹, R¹⁰, and R¹¹ are prepared as described supra.

[0243] The primary amines of formula I(h) may be reacted with theisocyanates or isothiocyantes of formula VII to prepare thecorresponding ureas and thioureas of formula I(k), see March, pages802-803.

[0244] Compounds of formula I where R is (CH₂)_(m′)CHR¹NHR², and R² isSO₂R⁷; and m, m′, R′, R¹, R⁷. R⁹, R¹⁰, and R¹¹ are prepared as describedsupra.

[0245] Compounds of formula I(f) may be converted to other compounds ofthe invention via standard combinatorial synthetic techniques. Forexample, a compound of formula I(f) dissolved or suspended in a suitablesolvent, optionally in the presence of a base, may be treated with acompound of formula VI to provide a compound of formula I(j) where R² isSO₂R⁷. Typically a preferred and convenient solvent is dichloromethane.When a base is employed, triethylamine is typically preferred.Furthermore, when a base is employed, the base and compound of formulaVI are typically employed in a slight stoichiometric excess. For examplea 1.01 to 1.40 molar excess, relative to the compound of formula I(f),is generally employed. About 1.15 to about 1.25 molar excess istypically preferred. When a base is not employed, the compound offormula VI is typically employed in a relatively larger stoichiometricexcess. For example, about a 1.5 to about a 3 molar excess, relative tothe compound of formula I(f), is usually employed. About 1.8 to about2.2 molar excess is typically prefer The reaction is usually performedat a temperature range of about 0° C. to about the reflux temperature ofthe solvent for from 10 minutes to 18 hours. Preferably, the reaction isperformed at about 15° C. to about 40° C. for from 5 minutes to about 1hour.

[0246] Compounds of formula I where R is (CH₂)_(m′)COR³ and R³ is C₁-C₆alkoxy, an amino acid ester, or NR⁴R⁵ may be prepared from compounds offormula I(i) as illustrated in Scheme 9 below where R¹⁴ is NR⁴R⁵, anamino acid ester, or C₁-C₆ alkoxy, R¹³ is an activating group, and m,m′, R′, R^(a), R^(b), R⁴, R⁵, R⁹, R¹⁰, and R¹¹ are as described supra.

[0247] Compounds of formula I(l), prepared as described in Scheme 1, mayalso be converted to other compounds of the invention via solution orsolid phase synthetic techniques. For example, acids of formula I(l) maybe treated with activating agents to form the activated carboxylic acidderivatives of formula VIII by methods well known in the chemical arts.See, e.g., The Peptides, Peptide Synthesis and the Examples andPreparations sections below.

[0248] Generally, preparation of compounds of formula I(m) where R¹⁴ isNR⁴R⁵ or an amino acid ester is performed in a manner similar to thereaction of compounds of formula I(f) or I(h) described in Scheme 6.Specifically, such compounds of formula I(m) may be prepared bydissolving or suspending a compound of formula VIII in a suitablesolvent, optionally in the presence of a suitable base, and adding anamine of formula m or IX. Typically a preferred and convenient solventis dichloromethane. Preferred bases are triethylamine andpiperidinylmethylpolystyrene resin. The amine is typically employed in amolar excess. For example, about a 1.5 to about a 3 molar excess,relative to the compound of formula VIII is usually employed. About 1.8to about 2.2 molar excess is typically preferred. The reaction isusually performed in a temperature range of about 0° C. to about thereflux temperature of the solvent for from 10 minutes to 18 hours.Preferably, the reaction is performed at about 15° C. to about 40° C.for from 5 minutes to about 2.5 hours.

[0249] The compounds of formula I(m) where R¹⁴ is C₁-C₆ alkoxy may beprepared by methods very well known in the chemical arts. Forinstruction on the conversion of activated carboxylic acid derivativesto esters see, e.g., Larock at 978-979. Alternatively, these compoundsof formula I(m) may be prepared directly from the acids of formula I(l)as taught in the Larock reference at pages 966-972.

[0250] The starting materials and compounds of the present invention maybe obtained by a number of routes. For example, compounds of formula IImay be prepared according to the routes shown in Schemes 8 and 9.

[0251] Where het, R^(a), R^(b), R, R′, R⁹, R¹⁰, and R¹¹, and m are asdescribed supra, compounds of formula II may be prepared according toScheme 10.

[0252] Compounds of formula II may be prepared by dissolving orsuspending a compound of formula XII in a suitable solvent and adding acompound of formula XI in a suitable solvent. Dimethylformamide is aconvenient solvent and is typically preferred for the compound offormula XII. A 1:1 mixture of DMF and dichloromethane is a convenientsolvent and is typically preferred for the amine of formula XI. Thisamide forming reaction is also preferably run in the presence ofdimethylaminopyridine (DMAP).

[0253] For compounds in which het is pyrazole, the addition of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) tothe reaction is preferred. The compound of formula XII is preferably thecorresponding carboxylic acid and is employed in an equimolar amount,relative to the compound of formula XI, but a slight excess (about a0.05 to about 0.15 molar excess) is acceptable. The DMAP is employed ina catalytic fashion. For example, about 5 molar percent to about 15molar percent, relative to the compound of formula A, is typicallyemployed. A 10 molar percent is usually preferred.

[0254] Compounds of formula XI where R is (CH₂)_(m′)COR¹,(CH₂)_(m′)NHPg, O(CH₂)₂NH-Pg, or (CH₂)_(m′)CO₂(C₁-C₆ alkyl) which areused to prepare compounds of formula I(d), I(f), I(h), and I(l)respectively, are well known in the art and to the extent notcommercially available, are readily synthesized by standard procedurescommonly employed in the art. Furthermore, the transformations describedin Schemes 3-8 may be performed before the cyclization described inScheme 1 and 2 to provide the compounds of formula XI with a fullyelaborated R substituent.

[0255] Additionally, compounds of formula XII may be prepared accordingto the following routes where, unless otherwise provided, R^(b) is thesubstituent from the carbon atom of het, R^(a) is the substituent fromthe saturated nitrogen of het, the second heteroatom is an oxygen orsulfur (represented by O in the following routes) and other variablesare as described supra.

[0256] Compounds of formula XI(a) may be prepared in a manner similar tothat described in the literature, for example, see Chen Y P, et. al,Heterocycles, 1995, 41, 175, and Chantegrel B, et. al, J. Org. Chem,1984, 49, 4419-4424.

[0257] Compounds of formula XVI may be prepared by dissolving orsuspending a compound of formula XV and a suitable base in a suitablesolvent and adding a compound of formula XIV in a suitable solvent,dropwise. Toluene is a convenient solvent and is typically preferred.Triethylamine is the preferred base. The compound of formula XIV istypically and preferably employed in an equimolar amount, relative tothe compound of formula XV, but a slight excess is acceptable.

[0258] The reactants are preferably combined at about 0° C. and theresulting solution is typically warmed to room temperature and mixed forfrom about 18 hours to about 24 hours.

[0259] The compound of formula XVI may then be converted to the compoundof formula XIII by dissolving or suspending a compound of formula XVI ina suitable acidic solvent and adding hydroxylamine hydrochloride.Glacial acetic acid is a convenient acidic solvent and is typicallypreferred. The ester group is then hydrolyzed to the correspondingcarboxylic acid of formula XI(a) through standard procedures commonlyemployed in the art.

[0260] The reactants are preferably combined at about room temperaturethen heated to reflux for from about 30 minutes to about 60 minutes.Preferably the action is heated to reflux from about 40 to 45 minutes.

[0261] The ester group of the compounds of formula XIII is thenhydrolyzed to the corresponding carboxylic acid through standardprocedures commonly employed in the art, see for example, Larock, pgs981-985.

[0262] Compounds of formula XIV and XV are known in the art and, to theextent not commercially available, are readily synthesized by standardprocedures commonly employed in the art.

[0263] Compounds of formula XIX may be prepared in a manner similar tothat described in the literature, for example, see Liu K, Shelton B R,Howe, R K, J. Org. Chem. 1980, 45, 3916-3918.

[0264] Generically, the compound of formula XVIII and hydroxylaminehydrochloride are suspended or dissolved in a suitable solvent and asuitable base is added. After the reaction is complete, the solution isacidified with a suitable acid and the resulting oxime is purified byknown methods. Typically a preferred and convenient solvent iswater/methanol. Typically a preferred and convenient base is sodiumhydroxide.

[0265] The reactants are preferably combined at about 0° C. and theresulting solution is typically mixed for about 1 hour at about 25-30°C., until the reaction is complete. After the reaction is complete, thesolution is acidified with a suitable acid, preferably hydrochloricacid, and the resulting oxime is purified by known methods.

[0266] The purified oxime is dissolved or suspended in a suitablesolvent, preferably DMF, and is then reacted with N-chlorosuccinimide(NCS). Preferably, NCS is added in small portions to control theexpected exotherm. The initial NCS addition results in a slighttemperature decrease. If the reaction does not self-initiate withinabout 10 minutes, as indicated by a slight temperature rise, hydrogenchloride may be bubbled into the DMF solution. If the reaction does notbegin within about 10 to 15 minutes, heating the solution to about45-60° C. is desirable. Once the reaction begins, the temperature ispreferably maintained below about 35° C. for benzaldoximes withelectron-donating substituents and below about 50° C. for strongelectron-withdrawing substituents. Completion of the reaction isindicated by cessation of the exotherm.

[0267] The compound of formula XIX is then converted to the compound offormula XX by methods well known to the skilled artisan. The compound offormula XIX and an appropriate methyl-2-butynoate are dissolved orsuspended in a suitable solvent, preferably ethyl ether, and Et₃N isadded. The reactants are combined at about room temperature and mixedfrom about 12 to 24 hours, until the reaction is complete.

[0268] The ester group of the compounds of formula XX is then hydrolyzedto the corresponding carboxylic acid of formula XI(b) through standardprocedures commonly employed in the art, see for example, Larock, pgs981-985.

[0269] Compounds of formula XU may be prepared in a manner similar tothat described in the literature, for example, see Stevens R L, AlbizatiK F, Tetrahedron Lett. 1984, 25, 4587. For an example of thistransformation, see Preparation 3.

[0270] Compounds of formula XI(c) may be prepared from compounds offormula Xe in a manner similar to that described in the literature, forexample, see Micetich R G, Chin C G, Can. J. Chem. 1970, 48, 1371. Foran example of this transformation, see Preparation 4.

[0271] Compounds of formula XVIII are known in the art and, to theextent not commercially available, are readily synthesized by standardprocedures commonly employed in the art.

[0272] The isoxazole compounds of formula XI may be converted to theisothiazole by methods well known in the art, for example see McGregor DN, Corbin U, Swigor J E, and Cheney L C, “Synthesis of isothiazoles: Thetransformation of isoxazoles into isothiazoles,” Tetrahedron, 1969, 25,389-395.

[0273] Compounds of formula I where het is pyrazolyl may be prepared asillustrated below.

[0274] Compounds of formula XXIII may be prepared by combining thecompound of formula XXII with ethyl trifluoroacetyl vinyl ether in amanner similar to that described in the literature, for example, seeKamitori et al, J. Het. Chem., 1993, 30, 389. For examples of thistransformation, see Preparations 7-8.

[0275] Additionally, compounds of formula XXIII may be prepared fromaldehyde hydrazones and ethyl propiolate as is further described byKamitori et al, Heterocycles, 1994, 38, 21.

[0276] The trifluoromethyl ketone of formula XXIII may be converted tothe corresponding carboxylic acid of formula XI(d) in a manner similarto that described in the literature, for example, see Delgado A, ClardyJ, Tetrahedron Lett. 1992, 33, 2789-2790.

[0277] Compounds of formula XXII are known in the art and, to the extentnot commercially available, are readily synthesized by standardprocedures commonly employed in the art.

[0278] Compounds of the invention where het is oxazolyl or imidazolylmay be prepared as illustrated below.

[0279] Compounds of formula XI(e) may be prepared in a manner similar tothat described in the literature, for example, see Nunami et al, J. Org.Chem. 1994, 59, 7635-7642.

[0280] The compound of formula XXV is converted to the compound offormula XXVIII by methods well known to the skilled artisan. Thecompound of formula XXV is dissolved or suspended in a suitable solvent,preferably tetrahydrofuran, and a base is added, preferably Et₃N. Thereactants are combined at about room temperature, preferably under aninert atmosphere, and mixed from about 30 minutes to 2 hours, until thereaction is complete.

[0281] The ester group of the compounds of formula XXVIII is thenhydrolyzed to the corresponding carboxylic acid formula XI(f) throughstandard procedures commonly employed in the art, see for example,Larock, pgs 981-985.

[0282] Compounds of formula XIV and XXIV are known in the art and, tothe extent not commercially available, are readily synthesized bystandard procedures commonly employed in the art.

[0283] The pharmaceutical salts of the invention are typically formed byreacting a compound of formula I with an equimolar or excess amount ofacid or base. The reactants are generally combined in a mutual solventsuch as diethylether, tetrahydrofuran, methanol, ethanol, isopropanol,benzene, and the like for acid addition salts, or water, an alcohol or achlorinated solvent such as dichloromethane for base addition salts. Thesalts normally precipitate out of solution within about one hour toabout ten days and can be isolated by filtration or other conventionalmethods.

[0284] Acids commonly employed to form pharmaceutical acid additionsalts are inorganic acids such as hydrochloric acid, hydrobromic acid,hydroiodic acid, sulfuric acid, phosphoric acid, and the like, andorganic acids such as p-toluenesulfonic, methanesulfonic acid,ethanesulfonic acid, oxalic acid, p-bromophenylsulfonic acid, carbonicacid, succinic acid, citric acid, tartaric acid, benzoic acid, aceticacid, and the like. Preferred pharmaceutical acid addition salts arethose formed with mineral acids such as hydrochloric acid, hydrobromicacid, and sulfuric acid, and those formed with organic acids such asmaleic acid, tartaric acid, and methanesulfonic acid.

[0285] Bases commonly employed to form pharmaceutical base additionsalts are inorganic bases, such as ammonium or alkali or alkaline earthmetal hydroxides, carbonates, bicarbonates, and the like. Such basesuseful in preparing the salts of this invention thus include sodiumhydroxide, potassium hydroxide, ammonium hydroxide, potassium carbonate,sodium carbonate, sodium bicarbonate, potassium bicarbonate, calciumhydroxide, calcium carbonate, and the like. The potassium and sodiumsalt forms are particularly preferred.

[0286] It should be recognized that the particular counterion forming apart of any salt of this invention is not of a critical nature, so longas the salt as a whole is pharmacologically acceptable and as long asthe counterion does not contribute undesired qualities to the salt as awhole.

[0287] The optimal time for performing the reactions of Schemes 1-8 andRoutes 1-5 can be determined by monitoring the progress of the reactionvia conventional chromatographic techniques. Furthermore, it ispreferred to conduct the reactions of the invention under an inertatmosphere, such as, for example, argon, or, particularly, nitrogen.Choice of solvent is generally not critical so long as the solventemployed is inert to the ongoing reaction and sufficiently solubilizesthe reactants to effect the desired reaction. The compounds arepreferably isolated and purified before their use in subsequentreactions. Some compounds may crystallize out of the reaction solutionduring their formation and then collected by filtration, or the reactionsolvent may be removed by extraction, evaporation, or decantation. Theintermediates and final products of formula I may be further purified,if desired by common techniques such as recrystallization orchromatography over solid supports such as silica gel or alumina.

[0288] The skilled artisan will appreciate that not all substituents arecompatible with all reaction conditions. These compounds may beprotected or modified at a convenient point in the synthesis by methodswell known in the art.

[0289] The following Preparations and Examples are provided to betterelucidate the practice of the present invention and should not beinterpreted in any way as to limit the scope of same. Those skilled inthe art will recognize that various modifications may be made while notdeparting from the spirit and scope of the invention. All publicationsmentioned in the specification are indicative of the level of thoseskilled in the art to which this invention pertains. The terms andabbreviations used in the instant Preparations and Examples have theirnormal meanings unless otherwise designated. For example “° C.”, “N”,“mmol”, “g”, “mL”, “M”, “HPLC”, “IR”, “MS(FD)”, “MS(IS)”, “MS(FIA)”,“MS(FAB)”, “MS(EI)”, “MS(ES)”, “UV”, and “¹H NMR”, refer to degreesCelsius, normal or normality, millimole or millimoles, gram or grams,milliliter or milliliters, molar or molarity, high performance liquidchromatography, infra red spectrometry, field desorption massspectrometry, ion spray mass spectrometry, flow injection analysis massspectrometry, fast atom bombardment mass spectrometry, electron impactmass spectrometry, electron spray mass spectrometry, ultravioletspectrometry, and proton nuclear magnetic resonance spectrometryrespectively. In addition, the absorption maxima listed for the IRspectra are only those of interest and not all of the maxima observed.

Preparations Preparation 1 2-Chloro-5-fluorobenzaldehyde Oxime

[0290]

[0291] To a mixture of 2-chloro-6-fluorobenzaldehyde (20 g, 0.126 mmol)in water (30 mL), ethanol (35 mL) and ice (55 g) were added tohydroxylamine hydrochloride (9.64 g, 0.139 mmol), then NaOH (12.6 g) inwater (30 mL) was added with stirring. Enough ice was added to maintaintemperature at 25-30° C. After 1 h, the mixture was washed with ether(150 mL) and acidified with concentrated HCl to a pH of 6 (ice was addedto keep the temperature at 25-30° C.). The product was extracted withdichloromethane (2×200 mL), dried over MgSO₄ and the solvent removed invacuo to furnish the oxime as a white solid (17.2 g, 79.0%). mp132.8-134.0° C.

[0292]¹H NMR (DMSO-d₆, 300 MHz) δ; 11.83 (s, 1H, OH), 8.19 (s, 1H,C:CH), 7.26-7.39 (m, 3H) ppm.

[0293] IR (CHC)₃) ν_(max) 3573, 3311, 1603, 1570, 1461, 1449, 1252, 986,964, 907, 845 cm⁻¹.

[0294] UV (EtOH) λ_(max) 249 (ε=11551) nm.

[0295] MS(ES) m/z 174.1 [(M+1)⁺, 100].

[0296] Anal. Calcd. for C₇H₅NOClF

[0297] Theoretical: C, 48.44; H, 2.90; N, 8.07, Cl, 20.43;

[0298] Found: C, 48.46; H, 2.91; N, 8.14, Cl, 20.18%.

Preparation 2 2-Chloro-5-fluorobenzoylchloride Oxime

[0299]

[0300] To a solution of alcohol (2 g, 11.53 mmol) in DMF (20 mL) wasadded N-chlorosuccinimide (NCS) (300 mg, 2.24 mmol) at room temperature.After 10 minutes, bubbled 10 mL HCl gas by syringe. When reactionstarted, cooled at −50° C. Slowly added rest of NCS (1.55 g, 11.61mmol). After two hours, the reaction mixture was poured into fourvolumes (80 mL) of ice water and extracted with ether (50 mL×2). Thecombined ether extracts were washed with water (20 mL×2), dried,concentrated to give the product as an oil (2.35 g, 98%). Because thiscompound was unstable, next reaction was carried out immediately.

[0301]¹H NMR (CDCl₃, 300 MHz) δ; 8.31 (s, 1H, OH), 7.37 (m, 1H), 7.27(d, J=7.7 Hz, 1H), 7.12 (t, 1H) ppm.

[0302] IR (CHCl₃) ν_(max) 3554, 3279, 1629, 1600, 1575, 1452, 1257,1230, 1180, 993, 939, 898 cm⁻¹.

[0303] UV (EtOH) λ_(max) 295 (ε=1359), 260 (ε=9105), 213 (ε=18662) nm.

Preparation 3 5-Chloro-3-(2-chloro-6-fluorophenyl)isoxazole

[0304]

[0305] A mixture of triethyl amine (44 mL) in DMF (50 mL) and vinylidenechloride (30 mL) was slowly added to a solution of methanol (26 g, 125mmol) in vinylidene chloride (100 mL) and DMF (100 mL) at roomtemperature under argon. After 15 h, the reaction mixture was pouredinto ether (700 mL) and washed with water (250 mL×3), dried, andconcentrated in vacuo to give crude product, which was purified by flashchromatography (gradient: 0-20% ethyl acetate/hexanes) to produced anoil as a product (23.3 g, 80.3%)

[0306]¹H NMR (CDCl₃, 300 MHz) δ; 7.31-7.44 (m, 2H), 7.13 (t, 1H), 6.40(s, 1H) ppm.

[0307]¹³C NMR (DMSO-d₆) δ 162.6 (C), 158.6 (C), 157.5 (C), 154.8 (C),143.6 (C), 131.7 (CH), 125.8 (CH), 114.5 (CH), 103.2 (CH) ppm.

[0308] IR (CHCl₃) ν_(max) 1612, 1560, 1454, 1392, 1253, 900 cm⁻¹.

[0309] UV (EtOH) λ_(max) 275 (ε=1103) nm.

[0310] MS (FIA) m/z 232.1 (M⁺, 100).

[0311] Anal. Calcd. for C₉H₄NOCl₂F

[0312] Theoretical: C, 46.59; H, 1.74; N, 6.04; Cl, 30.56;

[0313] Found: C, 46.45; H, 1.94; N, 6.15; Cl, 30.40%.

Preparation 4 4-Carboxy-5-chloro-3-(2-chloro-6-fluorophenyl)isoxazole

[0314]

[0315] Butyllithium (43.2 mL, 68.4 mmol) was slowly added to a solutionof chloroisoxazolic acid (13.8 g, 59.5 mmol) in THF (250 mL) at −78° C.The mixture was stirred for 30 minutes and carbon dioxide gas wasbubbled through the solution for 10 minutes. The solution was allowed towarm to room temperature in time hours and solvent was removed by vacuoto give a yellow salt which was washed with dry ether (100 mL×2).Addition of ethyl acetate (300 mL) dissolved the salt and concentratedHCl was dropped to the solution until a pH of 2. The product was washedwith water (50 mL), dried, and concentrated under the vacuo to givecrude product (10.0 g, 91.7%).

[0316] mp 163-164° C.

[0317]¹H NMR (CDCl₃, 300 MHz) δ; 9.61 (s, 1H, COOH), 7.42 (m, 1H), 7.29(t, 1H), 7.10 (t, 1H) ppm.

[0318] IR (CHCl₃) ν_(max) 1706, 1613, 1577, 1454, 1422, 1255, 1153,1147, 902 cm⁻¹.

[0319] UV (EtOH) λ_(max) 269 (ε=1505) nm.

[0320] MS(FIA) m/z 232.1 (M⁺, 100).

[0321] Anal. Calcd for C₁₀H₄NO₃Cl₂F

[0322] Theoretical: C, 43.51; H, 1.46; N, 5.07;

[0323] Found: C, 43.99; H. 1.40; N, 5.07%.

Preparation 5N-[3,4,5-trimethoxyphenyl)-3-(3-(2-chloro-5-fluorophenyl)-5-chloroisoxazol-4-oyl)aminophenyl-acetamide

[0324]

[0325] Thionyl chloride (5 mL) was added to a solution of4-carboxy-5-chloro-3-(2-chloro-6-fluorophenyl)isoxazole (130 mg, 0.471mmol) in dry toluene (10 mL). The resulting reaction mixture was stirredat reflux temperature for 4 h under nitrogen and then cooled to roomtemperature. The solvent was evaporated in vacuo and trace amounts ofthionyl chloride removed by coevaporation with toluene (2×25 mL).Resulting crude acid chloride (max. 0.471 mmol) was dissolved in THF (10mL) and a solution of amine (176 mg, 0.558 mmol) in THF:dichloromethane(1:1, 20 mL) was added dropwise and the mixture stirred at roomtemperature 6 h under nitrogen. The reaction mixture was evaporated todryness and the solid was titrated with 1N HCl (2×10 mL) to give a brownsolid which was further purified by flash chromatography (gradient:20-70% ethyl acetate/hexanes) to give product as a white solid (237 mg,89%).

[0326] mp 187-188° C.

[0327]¹H NMR (DMSO-d₆, 300 MHz) δ; 10.55 (s, 1H. NH), 10.08 (s, 1H, NH),7.37-7.61 (m, ArH₅), 7.25 (m, 1H, ArH), 7.04 (d, J=7.22 Hz, 1H), 6.93(s, ArH₂); 3.68 (s, 6H, 2OCH₃), 3.55 (s, 3H, OCH₃), 3.53 (s, 2H, CH₂)ppm.

[0328]¹³C NMR (DMSO-d₆) δ 168.7 (C), 162.1 (C), 158.1 (C), 157.0 (C),156.3 (C), 154.3 (C), 152.7 (C), 138.14 (C), 136.8 (C), 135.3 (C), 133.7(C), 133.6 (CH), 133.5 (CH), 132.1 (C), 128.8 (CH), 126.0 (CH), 125.9(Cl), 125.4 (CH), 120.5 (CH), 118.2 (CH), 115.3 (CH), 114.9 (CH), 114.0(C), 96.9 (CM), 60.1 (CH₃), 55.7 (CH₃), 43.4 (CH₂) ppm.

[0329] IR (CHCl₃) ν_(max) 1687, 1610, 1530, 1508, 1453, 1412, 1132 cm⁻¹.

[0330] UV (EtOH) λ_(max) 259 (ε=21847) nm.

[0331] MS(ES) m/z 574.1 (M⁺, 92).

[0332] Anal. Calcd. for C₂₇H₂₂N₃O₆Cl₂F

[0333] Theor.: C, 56.46; H, 3.86; N, 7.32; Cl, 12.32; F, 3.31;

[0334] Found: C, 56.34; H, 3.86; N, 7.04; Cl, 12.26; F, 3.39%.

Preparation 6N-(3,4,5-trimethoxyphenyl)3-(3-(2-chloro-5-fluorophenyl)-5-morpholino)isoxazol-4-oyl)aminopheny]acetamide

[0335]

[0336] To a solution of4-[3-(N-3,4,5-trimethoxy-benzylamidomethyl)phenyl-amido-5-chloro-3-(2-chloro-6-fluorophenyl)isoxazole(50 mg, 0.0871 mmol) in DMF (3 mL) was added morpholine (30.4 mg, 0.348mL). The mixture was stirred at room temperature for three and halfhours and then diluted with ethyl acetate (6 mL), washed with 1N HCl (6mL×3), dried over MgSO₄ and concentrated in vacuo to give crude productwhich was further purified by flash chromatography (gradient:hexanes/ethyl acetate, 8/2 to pure ethyl acetate) to give product as awhite solid (37.8 mg, 69.5%); Rf=0.37 (ethyl acetate/hexane, 713).

[0337]¹H NMR (CDCl₃. 300 MHz) δ; 6.76-7.43 (m, 6H), 6.74 (s, 2H), 3.87(m, 4H), 3.81 (m, 9H), 3.71 (m, 4H), 3.63 (s, 2H) ppm.

[0338] IR(CHCl₃) ν_(max) 1671, 1602, 1529, 1508, 1451, 1132 cm⁻¹.

[0339] UV (EtOH) λ_(max) 259 (ε=29874) nm.

[0340] MS(ES) m/z 625.3 (M⁺, 100).

[0341] Anal. Calcd. for C₃₋₁H₃₀N₄O₇ClF

[0342] Theoretical: C, 59.57; H, 4.84; N, 8.96; F, 3.04;

[0343] Found: C, 59.29; H, 5.13; N, 8.70; F, 3.30%.

Preparation 71-t-Butyl-3-(2-chloro-6-fluoro-phenyl)trifluoroacetyl-pyrazole

[0344]

[0345] 2-Chloro-5-fluorobenzaldehyde ten-butyl hydrazone hydrochloride(4.5 g, 0.017 mole) and sodium acetate (4.5 g, 0.025 mole) were taken ina 250 mL round bottom flask. Acetic acid (100 mL) and DMF (25 mL) wereadded to the flask and stirred for 5 minutes. Then, ethyltrifluoroacetyl vinyl ether (4.2 g) was added and stirred at 100° C. forone hour. The solvents were removed under vacuum and residue waschromatographed (silica column, 10% ethyl acetate in chloroform). Yieldwas 2.7 g, 46%.

[0346] NMR (CDCl₃): δ 1.70 (s, 9H, t-Bu), 7.00-7.15 (m, 1H, Ar),7.22-7.39 (m, 2K, Ar), 8.19 (s, 1H, pyr).

[0347] MS(ES): 349 (M⁺+1).

Preparation 81-t-Butyl-3-(2-chloro-6-fluorophenyl)-pyrazole-4-carboxylic Acid

[0348]

[0349] A compound from preparation 7 (2.8 g, 8 mmol) was dissolved inTHF (50 ml) and sodium hydride (1.5 g, 60% dispersion, 37 mmol) wasadded to the solution and stirred for a few minutes. Then water (0.5 ml)was added stirred at 60° C. for one hour. THF was removed under vacuum.The residue was dissolved in water and washed with ethyl acetate toremove any unreacted starting material. Then, the aqueous solution wasacidified with 5N HCl and extracted with ethyl acetate (3×100 mL). Thecombined ethyl acetate extracts were washed with brine, dried overNa₂SO₄, and evaporated to obtain the pyrazole acid. Yield: 1.7 g, 73%.

[0350] NMR (CDCl₃): δ 1.62 (s, 9H, t-Bu), 7.05 (t, 1H, Ar), 7.14 (d, 2H,Ar), 8.09 (s, 1H, pyr).

[0351] MS(ES): 297 (M+1)⁺, 295 (M−1)⁻.

Preparation 9N-(3,4,5-trimethoxyphenyl)3-(1-(tert-butyl)-3-(2-chloro-5-fluoro-phenyl)pyrazol-4-oyl)aminophenylAcetamide

[0352]

[0353] To a solution of a compound from preparation 8 (1 g, 3.37 mmol)dissolved in DMF (20 mL), EDCI (1.3 g, 6.74 mmol),2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (2.13 g, 6.75mmol) and DMAP (100 mg, catalyst) was added and stirred for 4 h at rt.The reaction mixture was diluted with ethylacetate (200 mL) and washedwith water (2×100 mL), acid (1 N HCl, 2×100 mL), brine (2×100 mL), driedover sodium sulfate, evaporated under vacuum and chromatographed (silicacolumn, 50% ethyl acetate in chloroform, isocratic). Yield 1.3 g, 65%.

[0354] NMR (CDCl₃): δ 1.65 (s, 9H, t-Bu), 3.62 (s, 2H, CH₂), 3.70 (s,3H, OMe), 3.71 (s, 6H, 2 OMe), 6.79 (s, 2H, Ar), 7.00-7.20 (m, 5H, Ar),7.20-7.50 (m, 2H, Ar), 8.23 (s, 1H, pyr).

[0355] MS(ES): 629 (M+34), 631 (M+36).

Preparation 10 3-Methyl-5-(2-chloro-6-fluorophenyl)₄-isoxazolecarboxylicAcid Ethyl Ester

[0356]

[0357] To a solution of ethyl-3-aminomethyl crotonate (4.79 g, 33.5mmol) in toluene (10 ml), was added triethylamine (3.73 g, 37 mmol). Thesolution was chilled using an ice bath, and then2-chloro-5-fluorobenzoyl chloride (6.47 g, 33.5 mmol) was added dropwiseover a 20 minute period. The reaction was allowed to warm slowly to roomtemperature, and stirred for 24 hours. The resulting suspension was thenfiltered, and the filtrate diluted with ethyl acetate (100 ml) andtransferred to a separatory funnel. The organic layer was sequentiallywashed with water, brine, dried (sodium sulfate), and the volatilesremoved under reduced pressure to provide the crude adduct (9.46 g) as agolden solid, and primarily one geometrical isomer.

[0358] NMR (CDCl₃) δ 6.95-7.4 (3 m, 3H), 3.8 (m, 2H, OEt), 3.12 (d, 3H,—NCH₃), 2.4 (s, 3H, vinyl CH₃), 0.8 (t, OEt). MS(ES) m/z 299.9 (M+H)⁺.

[0359] One major side product is acylation of the methylamino group.Crude adduct, however, was then redissolved in glacial HOAc (50 ml) towhich was then added NH₂OH.HCl (1.8 g, 1.1 eq). The solution was thenheated to reflux for 40-45 minutes to effect isoxazole formation. Thereaction mixture was concentrated to an oil, diluted with ether, andtransferred to a separatory funnel. The organic phase was washed withsaturated bicarbonate, brine, then dried. Filtration and concentrationafforded crude isoxazole ethyl ester (7.5 g), which could be usedwithout further purification.

[0360] NMR (CDCl₃) 8 (7.12, 7.3, 7.43, 3 m), 4.2 (q, 2H), 2.59 (s, 3H,CH₃), 1.1 (t, 3H).

[0361] MS(ES) m/z 283.9 (M+H)⁺.

Preparation 113-Methyl-5-(2-chloro-6-fluorophenyl)-4-isoxazolecarboxylic Acid

[0362]

[0363] Hydrolysis of the ethyl ester of3-methyl-5-(2-chloro-6-fluorophenyl)₄-isoxazolecarboxylic acid ethylester was accomplished by dissolving the crude ester (7.5 g, approx.0.027 mol) in THF (250 ml), and adding aqueous LiOH (1.344 g in 100 ml,2 eq). After stirring overnight at room temperature, the solution wasconcentrated to ⅔^(rd) volume, diluted with EtOAc (200 ml) and 50 mlwater, transferred to a separatory funnel, and the aqueous phasecollected. The organic phase was washed twice, and the combined aqueousphase was then acidified with 5N HCl. Back extraction with threewashings of EtOAc was then followed with a brine wash of the combinedorganics. After drying over Na₂SO₄, filtration and concentration, cleanisoxazole acid was obtained (2.94 g).

[0364] NMR (CDCl₃) δ 7.13, 7.32 7.46 (3m, 3H), 2.58 (s, CH₃).

[0365] MS(ES) m/z 253.8, 255.8 (M−W).

[0366] EA, calc'd for C₁₁H₇ClFNO₃

[0367] Theoretical: C, 51.68; H, 1.87; N, 5.48;

[0368] Found: C, 51.85; H, 1.87; N, 5.20%.

Preparation 12N-(3,4,5-Trimethoxyphenyl)3-(3-methyl-5-(2-chloro-5-fluoro-phenyl)isoxazol-4-oyl)aminophenylacetamide

[0369]

[0370] To 3-methyl-5-(2-chloro 5-fluorophenyl)₄-isoxazolecarboxylicacid, (38.4 mg, 0.15 mmol) in DMF (3 ml) was added DMAP (2 mg), followedby EDC.HCl (57 mg, 0.3 mmol). After a few minutes of stirring,2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (94.8 mg, 0.3mmol) was added, and the reaction proceeded for 18 hr at roomtemperature. The solution was then diluted with EtOAc (50 ml) and 1N HCl(20 ml), and transferred to a separatory funnel. The aqueous phase wasre-extracted twice with EtOAc, and the combined organics were thenwashed with saturated bicarbonate, brine, and dried. After filtration,the solution was concentrated to afford 20 mg of crude adduct. Thismaterial was purified by running the sample through a Bond-Elut Silicacartridge (500 mg) using 1:1 hexane/EtOAc as eluting solvent. Thus, apure sample (10 mg) of amide was obtained.

[0371] NMR (CDCl₃)δ 7.1, 7.2, 7.3, 7.4, 7.55 (5 m, 6H), 6.79 (s, 2H),3.8 (2s, 9H), 3.64 (s, 2H), 2.6 (s, 3H).

[0372] MS(ES) m/z 553.8 (M+H)⁺, 570.9 (M+NH₃)⁺.

[0373] EA, calc'd for C₂₈H₂₅ClFN₃O₆

[0374] Theoretical: C, 60.71; H, 4.55; N, 7.59;

[0375] Found: C, 59.83; H. 4.79; N, 6.83%.

Preparation 13 2-Fluorobenzaldehyde Oxime

[0376]

[0377] This reaction was carried out in a manner similar to thatdescribed in the literature (Liu, K. C.; Shelton, B. R.; Howe, R. K. J.Org. Chem. 1980, 45, 3916-3918). To a mixture of 2-fluorobenzaldehyde(5.20 g, 41.9 mmol) and hydroxylamine hydrochloride (3.20 g, 46.1 mmol),EtOH (15 ml), H₂O (15 ml), and ice (25 g) was added 50% NaOH (5.1 g in5.1 ml H₂O). The reaction was warmed to room temperature and stiffed for1 h. Ice was added to the reaction, followed by addition of 2N HCl untilthe pH was 2. The mixture was extracted with Et₂O. The organic extractwas washed (brine) and dried (MgSO₄). Filtration and concentration gaveoxime 2-fluorobenzaldehyde oxime (5.06 g, 87%) as a solid that was usedwithout further purification.

[0378]¹H NMR (300 MHz, CDCl₃) δ 9.02 (br S, 1H), 8.02 (m, 1H), 7.73 (m,1H), 7.35 (m, 1H), 7.20-7.00 (m, 2H) ppm.

[0379] Anal. calc. for C₇H₆FNO

[0380] Theoretical: C, 60.43; H, 4.35; N, 10.07;

[0381] Found: C, 60.55; H, 4.36; N, 10.24%.

[0382] MS(m/z) 140.2 [M+1].

Preparation 14 2-Fluorobenzoylchloride Oxime

[0383]

[0384] This reaction was carried out in a manner similar to thatdescribed in the literature (Liu, K. C.; Shelton, B. R.; Howe, R. K. J.Org. Chem. 1980, 45, 3916-3918). To a solution of 2-fluorobenaldehydeoxime (1.00 g, 7.2 mm l), in DMF (6 ml) was added N-chlorosuccinimide(0.99 g, 7.4 mmol). HCl (gas, 5 ml syringe taken from the vapors of aconcentrated solution of HCl) was bubbled into the solution. Afterstirring for 1 h, Et₂O and H₂O were added. The organic layer wasseparated, washed (H₂O and brine), and dried (MgSO₄). Filtration andconcentration gave 2-fluorobenzoylchloride oxime (0.96 g, 77%) as an offwhite solid that was used without further purification. ¹H NMR (300 MHz;CDCl₃) δ 9.18 (s, 1H), 7.71 (dt, 1H, J=1.5, 7.8 Hz), 7.44 (m, 1H), 7.23(app t, 1H, J=8.8 Hz), 7.16 (ddd, J=1.0, 8.3, 10.7 Hz) ppm.

[0385] Anal. calc. for C₇H₅ClFNO

[0386] Theoretical: C, 48.44; H, 2.90; N, 8.07;

[0387] Found: C, 48.80; H, 2.72; N, 8.04%.

[0388] MS(FD) (m/z) 174.0 [M+1].

Preparation 15 4-Methoxycarbonyl-5-methyl-3-(2-fluorophenyl)isoxazole

[0389]

[0390] To a solution (0° C., N₂) of 2-fluorobenzoylchloride oxime (0.50g, 2.9 mmol) and methyl-2-butynoate (0.57 g, 5.8 mmol) in Et₂O (15 ml)was added a solution of Et₃N (0.52 ml) in Et₂O (2 ml) over a period of 1h followed by warming to room temperature while stirring overnight (17h). The reaction was diluted with Et₂O, washed (H₂O then brine), dried(MgSO₄), filtered, and concentrated. Column chromatography (silica gel,hexanes/EtOAc gradient) gave the title compound (0.39 g, 58%).

[0391]¹H NMR (300 MHz, CDCl₃) δ 7.44 (m, 2H), 7.20 (m, 1H), 7;17 (m,1H), 3.68 (s, 3H), 2.70 (s, 3H) ppm.

[0392] MS(ES) (m/z) 236.0 [M+1].

Preparation 16 4-Carboxy-5-methyl-3-(2-fluorophenyl)isoxazole

[0393]

[0394] A solution of4-methoxycarbonyl-5-methyl-3-(2-fluorophenyl)isoxazole (2.20 g, 8.84mmol), MeOH (25 ml), and 2N NaOH (8.8 ml) was heated at 50° C. overnight(19 h). The reaction was cooled to room temperature, diluted with H₂O,and acidified (2N HCl) to less than pH 3. The mixture was extracted withEtOAc (twice) and the combined extracts were washed (brine), dried(MgSO₄), filtered, and concentrated to give the title compound (1.88 g,96%). This material was used without further purification.

[0395]¹H NMR (300 MHz, CDCl₃) δ 7.45 (m, 2H), 7.15 (m, 2H), 2.78 (s, 3H)ppm.

[0396] Anal. Calc. for C₁₁H₈FNO₃

[0397] Theoretical: C, 59.73; H, 3.65; N, 6.33;

[0398] Found: C, 59.38; H, 3.27; N, 6.85%.

[0399] MS(ES) (m/z) 220.2 [M−1].

Preparation 17N-(3,4,5-Trimethoxyphenyl)3-(5-methyl-3-(2-fluorophenyl)isoxazoloyl)aminophenylAcetamide

[0400]

[0401] To a solution of4-methoxycarbonyl-5-methyl-32-fluorophenyl)isoxazole (1.00 g, 4.52 mmol)in CH₂Cl₂ (30 ml) was added EDCI (1.51 g, 9.04 mmol), DMAP (0.11 g, 0.90mmol), and 2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (1.70g, 5.42 mmol). The reaction was allowed to stir for 6 h then applied toa silica gal column eluting first CH₂Cl₂ with followed by 10% acetone inCH₂Cl₂ to give the title compound (1.44 g, 62%).

[0402]¹H NMR (400 MHz, CDCl₃) δ 7.64-7.54 (m, 2H), 7.42 (br s, 1H), 7.36(dt, 1H, J=1.0, 7.3 Hz), 7.25 (m, 2H), 7.10 (m, 2H), 6.96 (d, 1H, J=7.8Hz), 6.75 (s, 2H), 3.80 (s, 6H), 3.78 (s, 3H), 3.64 (s, 2H), 2.78 (s,3H) ppm.

[0403] MS(ES) (m/z) 518.2 [M−1].

Preparation 18 2-Chloro-6-fluorobenzaldehyde Oxime

[0404]

[0405] To a solution of 2-chlorofluorobenzaldehyde (15.0 g, 94.6 mmol)and hydroxylamine hydrochloride (7.90 g, 114 mmol) in EtOH (25 ml), H₂O(25 ml), and ice (40 g) was added NaOH (19 ml, 50% wt. solution). Icewas added during the addition of the NaOH to maintain the reactiontemperature below 40 degrees C. The mixture was stirred at roomtemperature for 1 h. Ice was added, the pH adjusted to 6 with HCl (2 N),and extracted twice with Et₂O. The combined Et₂O layers were washed(brine) and dried (MgSO₄). Filtration and concentration gave ayellow-brown solid that was recrystallized from Et₂O/hexanes to give2-chloro-5-fluorobenzaldehyde oxime (12.2 g, 74%).

[0406]¹H NMR (300 MHz, CDCl₃) δ 9.82 (br s, 1H), 8.47 (s, 1H), 7.25 (m,2H), 7.07 (m, 1H) ppm.

[0407] Anal. calc. for C₇H₅ClFNO

[0408] Theoretical: C, 48.44; H, 2.90; N, 8.07;

[0409] Found: C, 48.73; H, 2.89; N, 7.95%.

[0410] MS(ES) (m/z) 174.1 [M+1].

Preparation 19 2-Chloro-6-fluorobenzoylchloride Oxime

[0411]

[0412] To a solution of 2-chloro 6-fluorobenzaldehyde oxime (5.13 g,29.6 mmol) in DMF (50 ml) was added a portion of N-chlorosuccinimidefollowed by a syringe of HCl gas (5 ml from the vapors of a conc. HClsolution). The temperature increased slightly and the rest of theN-chlorosuccinimide (total of 4.07 g, 30.5 mmol) was added. Afterstirring at room temperature for 1 h, ice was added and the mixture wasextracted twice with diethyl ether. The combined organic fractions werewashed (brine), dried (MgSO₄), filtered, and concentrated. Columnchromatography (silica gel, hexane/Et₂O gradient) gave2-chloro-6-fluorobenzoylchloride oxime (4.83 g, 78%).

Preparation 204-Methoxycarbonyl-5-phenyl-3-(2-chloro-6-fluorophenyl)isoxazole

[0413]

[0414] To a solution of 2-chlorofluorobenzoylchloride oxime (1.04 g,5.00 mmol) and methyl-3-phenyl-2-propynoate (1.20 g, 7.50 mmol) in Et₂O(25 ml) at 0° C. under nitrogen was added a solution of triethyl amine(0.834 ml, 6.00 mmol) in Et₂O (3 ml) over 1 h. The reaction was allowedto warm to room temperature overnight (17 h). The mixture was dilutedwith Et₂₀ and H₂O, the layers separated, and the organic fraction wasdried (MgSO₄). Column chromatography (silica gel, hexane/Et₂O gradient)gave the title compound (1.53 g, 92%).

[0415]¹H NMR (300 MHz, CDCl₃) g 8.05 (dd, 2H, J=2.0, 7.7 Hz), 7.55 (m,3H), 7.45-7.25 (m, 2H), 7.14 (t, 1H, H=8.3 Hz), 3.64 (s, 3H) ppm.

[0416] IR (CHCl₃ solution) ν 1725 (CO) cm⁻¹.

[0417] Anal. calc. for C₁₇H₁₁ClFNO₃

[0418] Theoretical: C, 61.55; H, 3.34; N, 4.22;

[0419] Found: C, 61.35; H, 3.42; N, 4.42%.

[0420] MS(ES) (m/z) 332.1 [M+1].

Preparation 21 4-Carboxy-5-phenyl-3-(2-chloro-6-fluorophenyl)isoxazole

[0421]

[0422] To a solution of4-methoxycarbonyl-5-phenyl-3-(2-chloro-4-fluorophenyl)isoxazole (0.70 g,2.1 mmol) in methanol (5 ml) and THF (2 ml) was added a solution of 2NNaOH (4 ml) at room temperature. The mixture was heated to 55° C. for1.5 h. Upon cooling to room temperature, aqueous HCl was added until thepH was below 4. The mixture was diluted with ethyl acetate and water andthe fractions separated. The organic fraction was washed (brine) anddried (MgSO₄). Filtration and concentration gave the title compound(0.67 g, 100%), which was used without further purification.

[0423] IR (KBr) ν 1696 (CO) cm⁻¹.

[0424] MS(ES) (m/z) 316.0 [M−1].

Preparation 22N-(3,4,5-trimethoxyphenyl)-3-(5-phenyl-3-(2-chloro-6-fluoro-phenyl)isoxazol-4-oyl)aminophenylAcetamide

[0425]

[0426] To a solution of4-carboxy-5-phenyl-3-(2-chloro-6-fluorophenyl)isoxazole (0.135 g, 0.425mmol) and 2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (0.129g, 0.408 mmol) in dichloromethane (5 ml) was added EDCI (0.082 g, 0.51mmol) and DMAP (0.062 g, 0.51 mmol) and stirred for 14 h. The mixturewas applied to a silica gel chromatography column. Elution with hexanesand ethyl acetate (gradient) gave the title compound (0.162 g, 64%).

[0427]¹H NMR (400 MHz, CDCl₃) δ 7.95 (dd, ¹H, J=1.7, 7.5 h), 7.57-7.50(m, 2H), 7.46-7.40 (m, 2H), 7.35-7.04 (m, 91), 6.73 (s, 2H), 3.79 (s,6H), 3.77 (s, 3H), 3.64 (s, 2H) ppm. MS(ES) (m/z) 616.2 [M+1].

Preparation 23 2,6-Difluorobenzaldehyde Oxime

[0428]

[0429] In a fashion similar to that for preparation 13,2,6-difluorobenzaldehyde (5.00 g, 35.2 mmol), hydroxylaminehydrochloride (2.93 g, 29.6 mmol), NaOH (7.04 g in 7.0 ml of water),ethanol (15 ml), water (15 ml), and ice (30 g) afforded2,6-difluorobenzaldehyde oxime (5.13 g, 93%).

[0430]¹H NMR (CDCl₃, 300 MHz) δ 9.85 (br s, 1H), 8.33 (s, 1H), 7.32 (m,1H), 6.95 (t, 2H, J=8.4 Hz) ppm.

[0431] MS(FD) (m/z) 157.0 [M+].

Preparation 24 2,6-Difluorobenzoylchloride Oxime

[0432]

[0433] In a fashion similar to that for preparation 14,2,6-difluorobenzaldehyde oxime (2.00 g, 12.7 mmol), N-chlorosuccinimide(1.75 g, 13.2 mmol), and DMF (20 ml) gave 2,6-difluorobenzoylchlorideoxime (1.58 g, 65%) after column chromatography (silica gel,hexanes/diethyl ether gradient).

[0434]¹H NMR (CDCl₃, 300 MHz) 58.56 (s, 1H), 7.55-7.37 (m, 1H), 6.95(app t, 2H, J=7.3 Hz) ppm.

[0435] MS(FD) (m/z) 191.0 [M+].

Preparation 254-Methoxycarbonyl-5-methyl-3-(2,6-difluorophenyl)isoxazole

[0436]

[0437] In a fashion similar to that for preparation 15,2,6-difluorobenzoylchloride oxime (1.00 g, 5.19 mmol),methyl-2-butynoate (1.01 g, 10.4 mmol), triethyl amine (0.94 ml, 6.8mmol) in diethyl ether (3 ml), and diethyl ether (25 ml) afforded4-methoxycarbonyl-5-methyl-3-(2,6-difluorophenylisoxazole (0.80 g, 61%)after purification by column chromatography (silica gel, hexanes/diethylether gradient).

[0438]¹H NMR (300 MHz, CDCl₃). δ 7.43 (m, 1H), 7.00 (app t, 2H, J=7.8Hz), 3.73 (s, 3H), 2.77 (s, 3H) ppm. IR (KBr) ν 1721 (CO) cm⁻¹.

[0439] MS(ES) (m/z) 254.1 [M+1].

[0440] Anal. Calc. for C₁₂H₉F₂NO₃

[0441] Theoretical: C, 56.92; H, 3.58; N, 5.53;

[0442] Found: C, 56.76; H, 3.48; N, 5.57%.

Preparation 26 4-Carboxy-5-methyl-3-(2,6-difluorophenyl)isoxazole

[0443]

[0444] In a manner similar to that for preparation 16,4-methoxycarbonyl-5-methyl-3-(2,6-difluorophenyl)isoxazole (0.244 g,0.910 mmol), 5N NaOH (1 ml, 5.0 mmol), and methanol (10 ml) afforded4-carboxy-5-methyl-3-(2,6-difluorophenyl)isoxazole (0.216 g, 93%) whichwas used without purification.

[0445]¹H NMR (400 MHz, CDCl₃) δ 7.39 (app tt, 1H, J=6.4, 8.3 Hz), 6.97(app t, 2H, J=8.3 Hz), 2.74 (s, 3H) ppm.

[0446] IR (CHCl₃ solution) ν 1698 (CO) cm⁻¹.

[0447] MS(ES) (m/z) 240.0 [M+1].

Preparation 27N-(3,4,5-Trimethoxyphenyl)-3-(5-methyl-3-(2,6-difluoro-phenyl)isoxazol-4-oyl)aminophenylAcetamide

[0448]

[0449] 4-Carboxy-5-methyl-32,6-difluorophenyl)isoxazole (0.120 g, 0.50mmol), 2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (0.174 g,0.55 mmol), EDCI (0.161 g, 1.00 mmol), DMAP (0.012 g, 0.10 mmol), anddichloromethane (5 ml) were allowed to react for 16 h in a fashionsimilar to that for example 5. Column chromatography (silica gel,hexanes/ethyl acetate gradient) afforded the title compound (0.142 g,53%).

[0450]¹H NMR (400 MHz, CDCl₃) δ 7.50 (app tt, 1H, J=5.0, 8.8 Hz), 7.39(br s, 1H), 7.25 (t, 1H, J=7.8 Hz), 7.10-7.02 (m, 5H), 6.98 (d, 1H,J=7.8 Hz), 6.71 (s, 2H), 3.77 (s, 6H), 3.74 (s, 3H), 2.77 (s, 3H).

[0451] MS(ES) (m/z) 538.2 [M+1].

Preparation 284-Methoxycarbonyl-5-hexyl-3-(2-chloro-6-fluorophenyl)isoxazole

[0452]

[0453] In a fashion similar to that for preparation 15,2-chloro-5-fluorobenzoylchloride oxime (0.750 g, 3.61 mmol),methyl-2-nonynoate (1.32 ml, 7.21 mmol), Et₃N (0.652 ml, 4.69 mmol) inEt₂O (3 ml), and Et₂O (15 ml) gave4-methoxycarbonyl-5-hexyl-3-(2,6-difluorophenyl)isoxazole (0.806 g, 66%)after column chromatography (silica gel, hexanes/Et₂O gradient).

[0454]¹H NMR (300 MHz, CDCl₃) d 7.39 (dt, 1H, J=5.9, 8.1 Hz), 7.29 (d,1H, J=8.1 Hz), 7.10 (dt, 1H, J=1.1, 8.4 Hz), 3.68 (s, 3H), 3.17 (t, 2H,J=7.7 Hz), 1.82 (m, 2H), 1.43-1.31 (m, 6H), 0.90 (m, 3H) ppm.

[0455] IR (CHCl₃ solution) ν 1723 (CO) cm⁻¹.

[0456] MS(ES) (m/z) 340.1 [M+1].

Preparation 29 4-Carboxy-5-hexyl-3-(2-chloro-6-fluorophenyl)isoxazole

[0457]

[0458] 4-Methoxycarbonyl-5-hexyl-3-(2,6-difluorophenyl)isoxazole (0.30g, 0.88 mmol) was dissolved in methanol (10 ml), 5N NaOH (1 nm) wasadded, and the mixture was heated at 50° C. for 2 h. Upon cooling toroom temperature, HCl was added until pH 1. Extraction with ethylacetate and subsequent washing of the organic fraction (brine), drying(MgSO₄), filtering, and concentrating afforded the desired4-carboxy-5-hexyl-3-(2,6-difluorophenyl)isoxazole (0.21 g, 72%) that wasused without further purification.

[0459]¹H NMR (400 MHz, CDCl₃) δ 7.34 (dt, 1H, 1=5.9, 8.3 Hz), 7.24 (d,1H, J=8.3 Hz), 7.05 (t, 1H, J=8.3 Hz), 3.13 (t, 2H, J=7.8 Hz), 1.78(quint, 2H, J=7.3 Hz), 1.40-1.25 (m, 6H), 0.85 (m, 3H) ppm.

[0460] IR (KBr) 3435 (br, OH), 1687 (CO) cm⁻¹.

[0461] MS(ES) (m/z) 324.4 [M−1].

Preparation 30N-(3,4,5-trimethoxyphenyl)-3-(5-hexyl-3-(2-chlorofluoro-phenyl)isoxazol-4-oyl)aminophenylAcetamide

[0462]

[0463] 4-Carboxy-5-hexyl-3-(2,6-difluorophenyl)isoxazole (150 mg, 0.46mmol), 2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (160 mg,0.51 mmol), EDCI (150 mg, 0.78 mmol), DMAP (12 mg, 0.10 mmol) werereacted in CH₂Cl₂ (5 ml) for 19 h then applied to a silica gel column.Elution with hexanes/EtOAc (gradient) gave the title compound (185 mg,65%).

[0464]¹H NMR (400 MHz, CDCl₃) δ 7.48 (dt, 1H, J=5.8, 8.3 Hz), 7.38 (d,1H, J=8.3 Hz), 7.34 (s, 1H), 7.26-7.18 (m, 2H), 7.03 (d, 1H, J=7.8 Hz),6.98 (s, 1H), 6.95 (s, 1H), 6.90 (d, J=9.3 Hz), 6.70 (s, 2H), 3.77 (s,6H), 3.75 (s, 3H), 3.61 (s, 2H), 3.19 (t, 2H, J=7.8 Hz), 1.82 (m, 2H),1.38 (m, 2H), 1.29 (m, 4H), 0.85 (m, 3H) ppm.

[0465] MS(ES) (m/z) 624.2 [M+1].

Preparation 31 2,4-Difluorobenzaldehyde Oxime

[0466]

[0467] In a fashion similar to that for preparation 13,2,4-difluorobenzaldehyde (5.00 g, 35.2 mmol), hydroxylaminehydrochloride (2.94 g, 42.3 mmol), NaOH (7.04 g, 88.0 mmol; as a 50%solution by weight in water), ethanol (15 ml), H₂O (15 ml), and ice (30g) gave 2,4-difluorobenzaldehyde oxime (4.62 g, 84%) which was usedwithout purification.

[0468]¹H NMR (300 MHz, CDCl₃) δ 8.26 (s, 1H), 7.66 (t, 1H, J=8.3 Hz),7.14-7.07 (m, 2H) ppm.

[0469] MS(EI) (m/z) 157.0 [M+].

Preparation 32 2,4-Difluorobenzoyl Chloride Oxime

[0470]

[0471] In a fashion similar to that for preparation 14,2,4-difluorobenzaldehyde oxime (2.52 g, 16.1 mmol), N-chlorosuccinimide(2.21 g, 16.5 mmol), and dimethyl formamide (25 ml) gave2,4-difluorobenzoyl chloride oxime (2.11 g, 68%) after chromatography(silica gel, hexanes/ether).

[0472]¹H NMR (400 MHz, CDCl₃) δ 8.48 (s, 1H), 7.62 (dt, 1H, J=6.4, 8.3Hz), 6.93-6.83 (m, 2H) ppm.

[0473] IR (CHCl₃ solution) ν 3555, 1613, 1506, 1429, 1272, 1258 cm⁻¹.

Preparation 33 4-Ethoxycarbonyl-5-methyl-3-(2,4-difluorophenyl)isoxazole

[0474]

[0475] In a fashion similar to that for preparation 15,2,4-difluorobenzoyl chloride oxime (2.00 g, 10.4 mmol), ethyl2-butynoate (1.76 g, 15.7 mmol), triethyl amine (2.9 ml, 21 mmol) indiethyl ether (10 ml), and diethyl ether 30 ml) gave4-ethoxycarbonyl-5-methyl-3-(2,4-difluorophenylisoxazole (1.62 g, 58%)after column chromatography (silica gel, hexanes/diethyl ethergradient).

[0476]¹H NMR (400 MHz, CDCl₃) δ 7.43 (1H, dt, J=6.4, 8.3 Hz), 6.93 (1H,app dt, J=2.9, 8.8 Hz), 6.86 (1H, app d, J≈2.4, 9.8 Hz), 4.16 (q, 2H,J=6.9 Hz), 2.70 (s, 3H), 1.14 (t, 3H, J=6.9 Hz) ppm.

[0477] IR (CHCl₃ solution) 1720 (CO) cm⁻¹.

[0478] MS(ES) (m/z) 268.3 [M+1].

Preparation 34 4-Carboxy-5-methyl-3-(2,4-difluorophenyl)isoxazole

[0479]

[0480] 4-Ethoxycarbonyl-5-methyl-3-(2,4-difluorophenyl)isoxazole (305mg, 1.14 mmol), 5N NaOH (1.0 ml, 5 mmol), methanol (5 ml), and THF (1ml) were heated at 50° C for 16 h. Ice was added and the pH was adjustedto 2-3 with 1N HCl. Extraction with EtOAc followed by washing (brine),drying (Na₂SO₄), filtering, and concentration gave the crude4-carboxy-5-methyl-3-(2,4-difluorophenyl)isoxazole (286 mg, 99%) whichwas used without further purification.

Preparation 35N-(3,4,5-trimethoxyphenyl)-3-(5-methyl-3-(2,4-difluoro-phenyl)isoxazol-4-oyl)aminophenylAcetamide

[0481]

[0482] 4-Carboxy-5-methyl-3-(2-difluorophenyl)isoxazole (97 mg, 0.41mmol) was dissolved (room temperature, N₂) in dichloromethane (5 ml)followed by addition of DMF (0.005 ml) and oxalyl chloride (0.070 i),0.82 mmol). After 1 h, the volatiles were removed. The residue and2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (110 mg, 0.35mmol) were dissolved in CH₂Cl₂ (5 ml) and DMF (1 ml) (room temperature,N₂), Et₃N (0.111 ml, 0.80 ml) and DMAP (5 mg, 0.04 mmol) were added. Thereaction was stirred for 3 h. The mixture was applied to a silica gelcolumn and elution (hexanes/EtOAc gradient) gave the title compound (152mg, 81%).

[0483]¹H NMR (400 MHz, CDCl₃) δ 7.60 (app q, 1H, J=7.8 Hz), 7.45 (s,1H), 7.28 (t, 1H, J=7.8 Hz), 7.20 (s, 1H), 7.10-7.00 (m, 5H), 6.97-6.92(m, 1H), 6.72 (s, 2H), 3.77 (s, 6H), 3.75 (s, 3H), 3.63 (s, 2H), 2.73(s, 3H) ppm.

[0484] MS(ES) (m/z) 538.2 [M+1].

Preparation 36 2-Fluoro-4-bromobenzaldehyde Oxime

[0485]

[0486] In a fashion similar to that for preparation 13,4-bromo-2-fluorobenzaldehyde (5.00 g, 24.6 mmol), hydroxylaminehydrochloride (2.05 g, 29.6 mmol), NaOH (4.92 g, 61.5 mmol; in a 50% byweight solution), ethanol (15 ml), water (15 ml), and ice (30 g) gavethe title compound (4.12 g, 77%) which was used without purification.

[0487]¹H NMR (400 MHz, CDCl₃) δ 8.26 (s, 1H), 7.57 (t, 1H, J=8.8 Hz),7.28-7.21 (m, 2H) ppm.

[0488] MS(ES) (m/z) 217.9 [M+1], 219.9 [M+3].

Preparation 37 2-Fluoro-4-bromobenzoyl Chloride Oxime

[0489]

[0490] In a fashion similar to that for preparation 14,2-Fluoro-4-bromo-benzaldehyde oxime (2.77 g, 12.7 mmol),N-chlorosuccinimide (1.76 g, 13.2 mmol), and DMF (25 ml) gave2-fluoro-4-bromo-benzoyl chloride oxime (2.61 g, 81%) after filtrationthrough a plug of silica gel eluting with hexanes/Et₂O.

[0491]¹H NMR (400 MHz, CDCl₃) δ 8.62 (s, 1H), 7.49 (dd, 1H, J=1.5, 7.8Hz), 7.34-7.29 (m, 2H) ppm.

[0492] Anal. calc. for C₇H₄BrClFNO

[0493] Theoretical: C, 33.30; H, 1.60; N, 5.55;

[0494] Found: C, 33.66; H, 1.58; N, 5.45%.

Preparation 384-Ethoxycarbonyl-5-methyl-3-(2-fluoro-4-bromophenyl)isoxazole

[0495]

[0496] In a fashion similar to that described for preparation 15,2-fluoro-4-bromo-benzoyl chloride oxime (2.00 g, 7.92 mmol),ethyl-2-butynoate (1.33 g, 11.9 mmol), triethylamine (2.2 ml, 15.8 mmol)in diethyl ether (10 ml), and diethyl ether (20 ml) gave4-ethoxycarbonyl-5-methyl-32-fluoro-4-bromophenyl)isoxazole (2.16 g,83%) after column chromatography (silica gel, hexanes/diethyl ethergradient). ¹H NMR (400 MHz, CDCl₃) δ 7.36-7.29 (m, 3H), 4.16 (q, 2HJ=6.8 Hz), 2.70 (s, 3H), 1.15 (t, 3H, J=6.9 Hz) ppm.

[0497] IR (KBr) ν 1719 (CO) cm⁻¹.

[0498] Anal. Calc. for C₁₃H₁₁BrFNO₃

[0499] Theoretical: C, 47.59; H, 3.38; N, 4.27;

[0500] Found C, 47.51; H, 3.30; N, 4.26%.

[0501] MS(ES) (m/z) 328.1 [M+1].

Preparation 39 4-Carboxy-5-methyl-3-(2-fluoro-4-bromophenyl)isoxazole

[0502]

[0503] In a fashion similar to that described for preparation 16 (120mg), 4-ethoxycarbonyl-5-methyl-3-(2-fluoro-4-bromophenyl)isoxazole (350mg, 1.06 mmol), 5N NaOH (1 ml, 5 mmol), methanol (5 ml), and THF (1 ml)gave 4-carboxy-5-methyl-3-(2-fluoro-4-bromophenyl)isoxazole (320 mg,100%) that was used without purification.

[0504] 1HNMR (300 MHz, CDCl₃) d 7.43-7.35 (m, 3H), 2.78 (s, 3H) ppm.

[0505] MS(EI) (m/z) 298.1[M−1], 300.1[M−3].

Preparation 40N-(3,4,5-Trimethoxyphenyl)-3-(5-methyl-3-(2-fluoro-4-bromophenyl)isoxazol-4-oyl)aminophenylacetamide

[0506]

[0507] In a fashion similar to that described for preparation 35,4-carboxy-5-methyl-3-(2-fluorobromophenyl)isoxazole (119 mg, 0.40 mmol),oxalyl chloride (0.070 ml, 0.80 mmol), dimethylformamide (0.010 ml),dichloromethane (5 ml),2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (110 mg, 0.35mmol), triethylamine (0.111 ml, 0.80 mmol), DMAP (5 mg),dimethylformamide (1 ml), dichloromethane (5 ml) gave the title compound(138 mg, 66%) after column chromatography (silica gel, hexanes/ethylacetate gradient).

[0508]¹H NMR (400 MHz, CDCl₃) δ 7.52-7.42 (m, 3H), 7.37 (dd, 1H, J=1.5,9.3 Hz), 7.29 (t, 1H, J=7.8 Hz), 7.20 (s, 1H), 7.09-7.04 (m, 4H), 6.72(s, 2H), 3.77 (s, 6H), 3.75 (s, 3H), 3.64 (s, 2H), 2.72 (s, 3H) ppm.

[0509] MS(ES) (m/z) 598.1 [M+1], 600.1 [M+3].

Preparation 414-Methoxycarbonyl-5-methyl(tetrahydropyran-2-yl)-3-(2-chloro-6-fluorophenyl)isoxazole

[0510]

[0511] 2-Chloro-4-fluoro-benzoyl chloride oxime (3.70 g, 17.8 mmol),alkyne 1 (for preparation see Earl, R. A. and Townsend, L. B. OrganicSynthesis 1981, 60, 81) (5.20 g, 26.7 mmol), triethyl amine (2.72 ml,19.6 mmol) in diethyl ether (3 ml), and diethyl ether (50 ml) werereacted in a fashion similar to that for example 3 to provide the titlecompound (4.32 g, 66%) after column chromatography (silica gel,hexanes/ethyl acetate gradient).

[0512]¹H NMR (400 MHz, CDCl₃) E 7.36 (dt, 1H, J=5.9, 8.3 Hz), 7.27 (m,1H), 7.07 (app t, 1H, J=8.9 Hz), 5.12 ({fraction (1/2)} AB, 1H, J=13.7Hz), 4.98 ({fraction (1/2)} AB, 1H, J=13.7 Hz), 4.84 (t, 1H, J=3.4 Hz),3.85 (m, 1H), 3.67 (s, 3H), 3.55 (m, 1H), 1.90-1.50 (m, 6H) ppm. IR(CHCl₃ solution) ν 1731 (CO) cm⁻¹.

[0513] Anal. calc. for C₁₇H₁₇ClFNO₅

[0514] Theoretical: C, 55.22; H, 4.63; N, 3.79;

[0515] Found: C, 55.24; H, 4.59; N, 3.78%.

Preparation 42N-(3,4,5-Trimethoxyphenyl)-3-(5-methyl(tetrahydropyran-2-yl)-3-(2-chloro-6-fluorophenyl)isoxazol-4-oyl)aminophenylAcetamide

[0516]

[0517] A mixture of4-methoxycarbonyl-5-methyl(tetrahydropyran-2-yl)-3-(2-chlorofluorophenyl)isoxazole(2.95 g, 7.98 mmol), 2N NaOH (8.0 ml, 16.0 mmol), MeOH (20 ml), and THF(10 ml) was heated to 55° C. for 4 h. After cooling to 0° C., HCl (1N)was added until pH 3. The mixture was extracted with ethyl acetate (3×),and the combined extracts were washed (brine), dried (MgSO₄), filtered,and concentrated to give the crude acid. The crude acid was dissolved indichloromethane (50 ml) along with2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (3.80 g, 12.0mmol), EDCI (3.33 g, 17.4 mmol), HOAt (2.72 g, 20.00 mmol), and DMAP(0.10 g, 0.80 mmol) and stirred at room temperature overnight. Themixture was applied to a silica gel column and eluted withdichloromethane/acetone (gradient) to give the title compound (2.61 g,41%).

[0518]¹H NMR (400 MHz, CDCl₃) δ 9.39 (s, 1H), 7.61 (s, 1H), 7.39-7.24(m, 4H), 7.09-7.04 (m, 2H), 6.69 (s, 2H), 5.00 (AB, 2H, J=14.2 Hz), 4.87(app t, 1H, J=3.0 Hz), 3.75 (s, 6H), 3.73 (s, 3H), 3.68 (m, 1H), 3.61(s, 2H), 3.55 (m, 1H), 1.80-1.40 (m, 6H) ppm.

[0519] MS(ES) (m/z) 652.3 [M−1].

Preparation 43 2-Iodo-6-fluorobenzaldehyde

[0520]

[0521] To a solution of diisopropylamine (6.94 ml, 49.5 mmol, 1.1 eq.)in THF (90 ml) under N₂, stirring at 0-5° C., was added dropwise over 10minutes n-BuLi (28.12 ml, 45 mmol) and stirred at this temperature for10 min. The reaction mixture was cooled to 78° C. in a dry ice/acetonebath and 1-iodo-5-fluorophenyl (5.29 ml, 45 mmol) was added dropwiseover 5 min. The reaction was stirred at this temperature for 1 h, thendropwise DMF (4.15 ml, 49.5 ml, 1.1 eq.) was added over 5 minutes, andstirred for 10 minutes. Acetic acid (9 ml) was added, followed by H₂O,and extracted with Et₂O. The combined organic solution was washed with0.1 N HCl, brine, dried (MgSO₄), filtered and concentrated. The crude2-iodo-6-fluorobenzaldehyde (9.65 g) was taken onto the next stepwithout purification. ¹H NMR (400 MHz, CDCl₃) δ 10.13 (s, 1H), 7.80 (d,1H), 7.31-7.21 (m, 2H).

[0522] GC[100 C (5 min.)→180 C (5 min.) @20 C/min], Rt=8.067.

Preparation 44 2-Iodo-6-fluorobenzaldehyde Oxime

[0523]

[0524] To a mixture of 2-iodo-6-fluorobenzaldehyde (45 mmol) in H₂O(22.5 ml), EtOH (22.5 ml), and ice (20 g) was added hydroxylaminehydrochloride (3.19 g, 49.5 mmol, 1.1 eq.). Then, 112.5 mmol of 50% NaOH(4.5 g in 4.5 ml H₂O, 2.5 eq.) was added with stirring. Enough ice tokeep the temperature at 25-30° C. was added. The reaction was stirredfor 2 h, acidified with conc. HCl to pH 4 (ice was added to keep thetemperature at 25-30° C.), and extracted with Et₂O. The combined organicsolution was washed with brine, dried (MgSO₄), filtered, andconcentrated. The residue was chromatographed on silica gel using ethylacetate in hexanes (5%-10%) to give 2-iodo-6-fluorobenzaldehyde oxime(7.06 g, 59% over 2 steps) as an oil that solidified over time.

[0525]¹H NMR (400 MHz, CDCl₃) δ 8.67 (br s, 1H), 8.29 (s, 1H), 7.71 (d,1H) 7.14 (t, 1H), 7.06-7.01 (m, 1H).

[0526] MS(ES) (m/z) 265.9 [M+1].

Preparation 45 2-Iodo-6-fluorobenzoyl Chloride Oxime

[0527]

[0528] To a stirred solution of 2-iodo-6-fluorobenzaldehyde oxime (7.06g, 26.6 mmol) in DMF (40 ml) at 25-30° C. was added about {fraction(1/5)} of 26.6 mmol (3.56 g) of NCS. The initial NCS addition results ina slight temperature decrease. If the reaction does not self-initiatewithin 10 min., as indicated by a slight temperature rise, 5 pipettevolumes of gas from the headspace of a conc. HCl reagent bottle isbubbled into the DMF solution. Careful addition of the rest of the NCSled to a temperature increase of 45-55° C. Once the reaction cools toR.T. (about 1 h), ice H₂O was added and the mixture was extracted withEt₂O. The combined organic solution was washed with brine, dried(MgSO₄), filtered, and concentrated. The crude 2-iodo-fluorobenzoylchloride oxime (8.1 g) was taken on to the next step withoutpurification.

[0529]¹H NMR (400 MHz, CDCl₃) δ 8.34 (s, 1H), 7.68 (d, 1H), 7.16-7.12(m, 2H).

[0530] IR (KBr) 3573.39, 3313.25, 1596.58, 1560.15, 1455.53, 1441.90,1250.77, 979.18, 956.22, 889.04, 820.56 cm⁻¹.

Preparation 464-Ethoxycarbonyl-5-methyl-3-(2-iodo-6-fluorophenyl)isoxazole

[0531]

[0532] To a solution of 2-iodo-6-fluorobenzoyl chloride oxime (26.6mmol) and ethyl-2-butynoate (3.72 ml, 31.92 ml, 1.2 eq.) in Et₂O (106ml) under N₂, stirring at 0-5° C., was added dropwise a solution of Et₃N(4.81 ml, 34.6 mmol, 1.3 eq) in Et₂₀ (13 ml) over 1 h. The reaction wasallowed to warm to room temperature and stirred overnight Et₂O was addedand then washed with H₂O, brine, dried (MgSO₄), filtered, andconcentrated. The crude residue was chromatographed on silica gel usingethyl acetate in hexanes (1-20%) to give the tide compound (8.21 g,82%).

[0533]¹H NMR (400 MHz, CDCl₃), 7.71 (d, 1H), 7.17-7.13 (m, 2H), 4.16 (q,2H), 2.78 (s, 3H), 1.05 (t, 3H).

[0534] IR (KBr) 1719.58, 1610.33, 1565.58, 1445.69, 1309.24, 1250.19,1130.60, 1104.46, 858.18 cm⁻¹.

Preparation 47 4-Carboxy-5-methyl-32-iodo-6-fluorophenyl)isoxazole

[0535]

[0536] To a solution of4-ethoxycarbonyl-5-methyl-3-(2-iodo-6-fluoro-phenyl)isoxazole (7.15 g,19 mmol) in EtOH (100 ml) was added 5N NaOH (7.6 ml, 38 mmol, 2 eq.),and stirred in a 50° C. oil bath for 4 hrs. The reaction was cooled toroom temperature, poured into ice H₂O and extracted with Et₂O. Theaqueous layer was then acidified to pH 3 using conc. HCl and extractedwith ethyl acetate. The organic solution was washed with brine, dried(MgSO₄), filtered, and concentrated to give the title compound (6.08 g,92%).

[0537]¹H NMR (400 MHz, CDCl₃) δ 7.71 (d, 1H), 7.17-7.16 (m, 2H), 2.79(s, 3H).

[0538] IR (KBr) 1698.15, 1606.56, 1567.35, 1446.24, 1250.53, 858.75cm⁻¹.

Preparation 48N-(3,4,5-Trimethoxyphenyl)₃-(5-methyl-32-iodofluorophenyl)isoxazol-4-oyl)aminophenylAcetamide

[0539]

[0540] To a solution of4-carboxy-5-methyl-342-iodo-6-fluoro-phenyl)isoxazole (1.75 g, 5 mmol)in dichloromethane (30 ml) and THF (1 ml) was added oxalyl chloride(0.88 ml, 10 mmol, 2 eq.) under N₂, then DMF (3 drops) to initiate thereaction. The reaction was stirred for 2 h, concentrated, andre-dissolved in dichloromethane (50 ml) under N₂. A solution of2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (1.59, 5 mmol) inTHF (14 ml) was added, cooled to 0° C., and then added triethylamine(0.7 ml, 5 mmol), and stirred for 17 h. Crude reaction was partitionedbetween ethyl acetate and 1N HCl. The organic layer was washed with H₂Oand brine, dried (MgSO₄), filtered and concentrated. The crude materialwas titrated with diethyl ether (3×) and purified by flashchromatography (silica gel) using 1-10% acetone in dichloromethane togive the title compound (1.67 g, 51%).

[0541] MS(ES) (m/z) 646.19 [M+1].

[0542] IR (KBr) 1706.18, 1666.12, 1608.97, 1561.18, 1548.78, 1507.10,1491.46, 1451.15, 1409.43, 1243.72, 1225.47, 1214.50, 1177.72, 1129.46,854.90, 779.41 cm⁻¹.

Preparation 49 2-Fluoro-5-iodobenzaldehyde

[0543]

[0544] To a solution of diisopropylamine (1.54 ml, 1 mmol, 1.1 eq.) inTHF (20 ml) under N₂ stirring at 0-5° C. was added dropwise over 10minutes 1.6M n-BuLi (6.25 ml, 100 mmol) and stirred at this temperaturefor 10 min. The reaction mixture was cooled to −78° C. in a dryice/acetone bath and dropwise added 1-iodofluorophenyl (1.12 ml, 10mmol) over 5 min. The reaction was stirred at this temperature for 1 h,and DMF (0.8 ml, 11 mmol, 1.1 eq.) was added dropwise over 5 minutes,and stirred for 10 minutes. Acetic acid (2 ml) was added, followed byH₂O and extracted with Et₂O. The combined organic solution was washedwith 0.1 N HCl, brine, dried (MgSO₄), filtered and concentrated. Thecrude 2-fluoro-5-iodobenzaldehyde (2.36 g) was taken onto the next stepwithout purification.

[0545]¹H NMR (400 MHz, CDCl₃) δ 10.26 (s, 1H), 8.14 (d, 1H), 7.89-7.86(m, 1H), 6.95 (t, 1H).

[0546] GC[100 C (5 min.)→180 C (5 min.) @20 Clmin], Rt=7.375.

Preparation 50 2-Fluoro-5-iodobenzaldehyde Oxime

[0547]

[0548] To a mixture of 2-fluoro-5-iodobenzaldehyde (9.4 mmol) in H₂O (5ml), EtOH (5 ml), and ice (4.5 g) was added hydroxylamine hydrochloride(0.67 g, 10.3 mmol, 1.1 eq.). Then, 23.6 mmol of 50% NaOH (1 g in 1 mlH₂O, 2.5 eq.) was added with stirring. Enough ice to keep thetemperature at 25-30° C. was added. The reaction was stirred for 2 h,acidified with conc. HCl to pH 4 (ice was added to keep the temperatureat 25-30° C.), and extracted with Et₂O. The combined organic solutionwas washed with brine, dried (MgSO₄), filtered, and concentrated. Theresidue was chromatographed on silica gel using (1-10%) ethyl acetate inhexanes to give 2-fluoro-5-iodobenzaldehyde oxime (2.06 g, 78% over 2steps).

[0549] MS(ES) (m/z) 264.0 [M−1].

[0550] IR (KBr) 3575.39, 1478.30, 1262.00, 1238.75, 1109.23, 964.08,810.03, 809.87 cm⁻¹.

Preparation 51 2-Fluoro-5-iodobenzoyl Chloride Oxime

[0551]

[0552] To a stirred solution of 2-fluoro-5-iodobenzaldehyde oxime (2.06g, 7.77 mmol) in DMF (10 ml) at 25-30° C. was added about 1/5 of 7.77mmol (1.04 g) of NCS. The initial NCS addition results in a slighttemperature decrease. If the reaction does not self-initiate within 10min., as indicated by a slight temperature rise, 5 pipette volumes ofgas from the headspace of a conc. HCl reagent bottle is bubbled into theDMF solution. The rest of the NCS was carefully added, and thetemperature rose to 3540° C. Once the reaction cooled to R.T. (about 2h), ice H₂O was added and the crude was extracted with Et₂O. Thecombined organic solution was washed with brine, dried (MgSO₄),filtered, and concentrated. The crude product 2-fluoro-5-iodobenzoylchloride oxime (2.24 g) was taken on to the next step withoutpurification.

[0553] MS(FD) (m/z) 298.9.

[0554] IR (KBr) 3553.03, 3258.13, 1601.51, 1484.38, 1396.69, 1266.05,1241.83, 1122.00, 1000.66, 954.27, 820.20 cm⁻¹.

Preparation 524-Ethoxycarbonyl-5-methyl-3-(2-fluoro-5-iodophenyl)isoxazole

[0555]

[0556] To a solution of 2-fluoro-5-iodobenzoyl chloride oxime (2.24 g,7.48 mmol) and ethyl-2-butynoate (1.74 ml, 14.96 mmol, 2 eq.) in Et₂₀(28 ml) under N₂ stirring at 05° C., was added dropwise a solution ofEt₃N (1.35 ml, 9.72 mmol, 1.3 eq) in Et₂₀ (3.6 ml) over 1 h. Reactionwas allowed to warm to room temperature and stirred overnight. It wasthen partitioned between Et₂O and H₂O, washed with brine, dried (MgSO₄),filtered, and concentrated. The crude residue was chromatographed onsilica gel using (1-10%) ethyl acetate in hexanes to give the titlecompound (2.06 g, 73%).

[0557] MS(ES) (m/z) 376.2 [M+1].

[0558] IR (KBr) 1719.10, 1606.15, 1454.85, 1311.13, 1261.44, 1233.84,1122.63, 1099.04, 819.26 cm⁻¹.

Preparation 53 4-Carboxy-5-methyl-3-(2-fluoro-5-iodophenyl)isoxazole

[0559]

[0560] To a solution of4-ethoxycarbonyl-5-methyl-3-(2-fluoro-5-iodophenyl)isoxazole (1.04 g,2.77 mmol) in EtOH (14 ml) was added 5N NaOH (1.11 ml, 5.6 mmol, 2 eq.)and stirred in a 50° C. oil bath for 1 hr. Rxn was cooled to roomtemperature, poured into ice H₂O and extracted with Et₂O. Acidifiedaqueous layer to pH 2 using conc. HCl and extracted with ethyl acetate.The combined organic solution was washed with brine, dried (MgSO₄),filtered, and concentrated to give the title compound (0.85 g, 86%).

[0561] MS(ES) (m/z) 348.0 [M+1].

[0562] IR (KBr) 1698.84, 1602.67, 1465.80, 1261.94, 1234.74, 819.32cm⁻¹.

Preparation 54N-(3,4,5-Trimethoxyphenyl)-3-(5-methyl-32-fluoro-5-iodo-phenyl)isoxazol-4-oyl)aminophenylAcetamide

[0563]

[0564] To a solution of4-carboxy-5-methyl-3-(2-fluoro-5-iodophenyl)isoxazole (0.85 g, 2.45mmol) in dichloromethane (15 ml) was added oxalyl chloride (0.5 ml, 4.9mmol, 2 eq.) under N₂, then DMF (3 drops) to catalyze the reaction.Reaction was stirred for 1 h, concentrated, and dissolved indichloromethane (25 ml) under N₂.2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide was added (0.77 g,2.45 mmol), reaction cooled to 0° C., added triethylamine (0.33 ml, 2.45mmol), and then stirred for 2 h 30 min. Crude was partitioned betweenethyl acetate and 1N HCl. Organic layer was washed with H₂O and brine,dried (MgSO₄), filtered and concentrated. The crude material wastriturated with Et₂O (3×) to give the title compound (1.0 g, 63%).

[0565] MS(ES) (m/z) 646.4 [M+1].

[0566] IR (KBr) 1681.31, 1605.86, 1508.01, 1452.11, 1132.40 cm⁻¹.

Preparation 553-[9-chloro-3-mesylmethyl-isoxazolo[4,5-c]-quinoline-4(5H)-one]-N-3,4,5-trimethoxyphenyl-benzeneAcetamide

[0567]

[0568] Under the conditions described for example 20, alcohol3-[9-chloro-3-methoxy-isoxazolo[4,5-c]-quinoline-4(5H)-one]-N-3,4,5-trimethoxyphenyl-benzeneacetamide (226 mg, 0.41 mmol), methanesulfonyl chloride (0.16 ml, 2.1mmol), triethyl amine (0.45 ml, 3.2 mmol), and dichloromethane (6 ml)gave the crude title compound (241 mg, 94%) which was used withoutfurther purification.

Preparation 56 4-Methoxycarbonyl-5-(2-chloro-6-fluorophenyl)oxazole

[0569]

[0570] To a solution of 2-chloro-6-fluorobenzoic acid (2.14 g, 12.3mmol), CH₂Cl₂ (50 ml), and DMF (2 drops) at room temperature under N₂was added oxalyl chloride (9.2 ml, 18.4 mmol) dropwise. The mixture wasstirred for 3 h and the volatiles were removed under reduced pressure togive the crude acid chloride. The crude acid chloride and methylisocyanoacetate (1.23 g, 12.3 mmol) were dissolved in THF (20 ml) atroom temperature under N₂ and Et₃N (6.8 ml, 49.2 mmol) was addeddropwise over a period of 30 minutes. The reaction was allowed to stirfor 22 h followed by dilution with Et₂O, extraction with H₂₀, 1N HCl,H₂O, brine, dried (MgSO₄), filtered, and concentrate. Columnchromatography (silica gel, hexanes/Et₂O gradient) gave4-methoxycarbonyl-5-(2-chloro-6-fluorophenyl)oxazole (0.74 g, 24%).

[0571]¹H NMR (400 MHz, CDCl₃) d 8.01 (s, 1H), 7.41 (dt, 1H, J=5.9, 8.3Hz), 7.29 (ap d, 1H, J=8.3 Hz), 7.09 (ap t, 1H, J=8.3 Hz), 3.81 (s, 3H)ppm.

Preparation 57N-(3,4,5-trimethoxyphenyl)-3-(5-(2-chlorofluoro-phenyl)oxazol-4-oyl)aminophenylacetamide

[0572]

[0573] A mixture of 4-methoxycarbonyl-5-(2-chloro-6-fluorophenyl)oxazole(236 mg, 0.927 mmol), MeOH (5 ml), THF (1 ml), and 2N NaOH (1.5 ml, 3.00mmol) was heated to 50° C. for 30 minutes. The mixture was cooled toroom temperature and H₂O and ice were added followed by acidification topH 2 and extraction with EtOAc. The organic fraction was washed (brine),dried (Na₂SO₄), filtered, and concentrated to give the crude carboxylicacid. The crude carboxylic acid,2-(3-aminophenyl)-N-(3,4,5-trimethoxyphenyl)acetamide (292 mg, 0.927mmol), EDCI (354 mg, 1.85 mmol), and DMAP (11 mg, 0.093 mmol) wereallowed to react in CH₂Cl₂ (5 ml) for 15 h at room temperature under N₂.The mixture was applied directly to a silica gel column and elution withhexanes/EtOAc (gradient) gave the title compound (193 mg, 39%).

[0574]¹H NMR (400 MHz, CDCl₃) d 8.88 (s, 1H), 8.02 (s, 1H), 7.72 (s,1H), 7.48 (d, 1H, J=8.3 Hz), 7.40 (dt, 1H, J=5.9, 7.3 Hz), 7.33-7.28 (m,2H), 7.12-7.03 (m, 2H), 6.68 (s, 2H), 3.75 (s, 6H), 3.74 (s, 3H), 3.64(s, 2H) ppm.

[0575] Mass spectrum (ES) (m/z) 540.1 [M+1].

Preparation 583-(2-Chloro-6-fluorophenyl)₅-methyl-4H-pyrazole-4-carboxylic Acid EthylEster

[0576]

[0577] 2-(2-Chloro-6-fluoro-benzoyl)-3-methylamino-but-2-enoic acidethyl ester (500 mg, 1.6 mmol) was taken in acetic acid (10 mL) and wastreated with hydrazine hydrate at 150° C. overnight. Acetic acid isremoved under reduced pressure and the residue was dissolved in ethylacetate (20 mL) and washed with water, (2×10 mL), brine (2×10 mL), driedover sodium sulfate, filtered and evaporated to yield the title compound(588 mg, crude).

[0578] MS(ES): 283 (M)⁺, 284 (M+1)⁺, 281 (M−1)⁻.

[0579] HNMR (CDCl₃): 1.03-1.06 (t, 3H), 2.55 (s, 3H), 7.03-7.07 (t, 1H),7.21-7.7.35 (m, 2H).

Preparation 59Methyl-2-(formylamino)-3-(2-chloro-6-fluorophenyl)acrylate

[0580] NaH (60% in oil, 2.4 g, 0.06 mol, approx. 1.2 eq) was placed in adry 50 ml flask and heated to 35 degrees. To it was slowly added 50 mlof a THF solution containing 2-chloro-6-fluorobenzaldehyde (7.93 g, 0.05mol) and methyl isocyanoacetate (5 g, 0.05 mol). The reaction was thencooled to RT, and stirred for 3 hr. At that time, 10 ml of 10% HOAc wasadded. After 10 min, the contents were concentrated in vacuo to a darkbrown oil, then diluted with CHCl₃, and transferred to a sep funnel. Theaqueous phase was reextracted (3×) with CHCl₃, and the combined extractswere washed with brine, dried over Na₂SO₄, filtered, and concentrated tocrude product. Purification over a pad of silica gel was accomplished byelution with 10:1 CHCl₃/EtOAc to afford the title compound, clean Z/Eolefins (1:1 mix, 8.24 g, 64%).

[0581] NMR (CDCl₃) δ 8.48 (s, vinyl H), 8.09 (s, vinyl H), 7.8 (brd s,HCO), 7.1 (m), 7.0 (m), 3.65 (s, E 3), 3.6 (Z-CH₃).

[0582] MS (+ES) 257.91259.9 (M+H)+.

Preparation 60Methyl-2-(formylamino)-3-bromo-3-(2-chloro-6-fluorophenyl)acrylate

[0583] To the chilled (0°) olefin mixture (3.723 g, 14.5 mmol) in CCl₄(40 ml) was added N-bromosuccinimide (3.874 g, 21.8 mmol) slowly over a5 min period. After 4 hr, the reaction was complete, and Et₃N (1.52 g,15 mmol) was added dropwise with vigorous stirring. The resultingmixture was diluted with EtOAc (250 ml, total) and transferred to aseparatory funnel. The organic phase was washed with saturated aqueousbicarbonate, then brine, and dried over Na₂SO₄. Following filtration andconcentration, the crude product (predominantly Z olefin) was obtained,and purified by filtration over a pad of silica gel (2:1 hex/EtOAc) toafford the title compound (2.4 g, 50%).

[0584] NMR (CDCl₃) δ 8.25 (brd s, HCO), 7.1-7.2 (m), 6.98 (app t), 3.5(s, CH₃).

[0585] MS (−ES) 333.8/335.8 (M−H)−.

Preparation 61Z-Methyl-2-isocyano-3-bromo-32-chloro-6-fluorophenyl)acrylate

[0586] To the olefin (1.2 g, 3.58 mmol) and Et₃N (0.896 g, 8.96 mmol) in10 ml MeCl₂ chilled to −200 was added POCl₃ (0.37 ml, 3.94 mmol)dropwise. The reaction mixture was then warmed to 00, and stirred for3.5 hr. At that time, the reaction was poured into a separatory funnelcontaining 20% aq. K₂CO₃ (50 ml) and MeCl₂ (50 ml). The basic aqueoussolution was then washed with additional organic solvent, and thecombined extracts were washed with brine, dried over Na₂SO₄, filtered,and concentrated to afford crude product. Purification (silica gelchrom., MeCl₂) afforded clean isonitrile (0.325 g, 45%).

[0587] NMR (CDCl₃) δ 7.0-7.3 (3 m), 3.72 (s, CH₃).

[0588] IR (CHCl₃) 2117 cm⁻¹.

EXAMPLES Example 1N-(3,4,5-Trimethoxyphenyl)3-(5-chloroisoxazol[3,4-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide

[0589]

[0590] To a solution ofN-[3,4,5-trimethoxyphenyl)-3-(3-(2-chloro-5-fluorophenyl)-5-chloroisoxazol-4-oyl)aminophenyl-acetamide(100 mg, 0.1742 mmol) in DMF (40 mL) was added potassium carbonate (70mg, 0.522 mmol) at 0° C. The mixture was stirred at −10° C. for threehours. The solution was allowed to warm to room temperature and stirredfor three more hours. The reaction mixture was diluted with ethylacetate (50 mL) and washed with 1N HCl (100, 80, 40 mL), dried andconcentrated in vacuo to give crude product which was further purifiedby flash chromatography (silica gel) (gradient: hexanes/ethyl acetate)to give product as a white solid (30 mg, 31.1%); Rf=0.68 (ethylacetate/hexane, 7/3).

[0591]¹H NMR (CDCl₃, 300 MHz) 8; 10.18 (s, 1H, NH), 7.38-7.69 (m, ArH₇),6.95 (s, ArH₂); 3.68 (s, 6H, 2OCH₃), 3.67 (s, 2H, CH₂), 3.58 (s, 3H,OCH₃) ppm.

Example 2N-(3,4,5-Trimethoxyphenyl)-3-(5-morpholin-4-ylisoxazolo[3,4-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide

[0592]

[0593] To a solution ofN-(3,4,5-trimethoxyphenyl)3-(3-(2-chloro-5-fluorophenyl)-5-morpholino)isoxazoloyl)aminophenylacetamide (140 mg, 0.224 mmol) in DMF (6.9 mL) was added potassiumcarbonate (247.5 mg, 1.79 mmol). The mixture was stirred at roomtemperature for four hours and then diluted with ethyl acetate (20 mL),washed with water (6 mL×3), dried over MgSO₄ and concentrated to givecrude product, which was further purified by flash chromatography togive product as a white solid (131 mg, 97%).

[0594] IR (CHCl₃) ν_(max) 3010, 1663, 1601, 1594, 1497, 1451, 1412,1263, 1132 cm⁻¹;

[0595] UV (EtOH) λ_(max) 298 (ε=16442), 289 (ε=15852), 256 (ε=34088),249 (ε=31992), 215 (ε=51994) nm.

[0596] MS (ES) m/z 605.3 (M⁺, 100);

[0597] Anal. Calcd. for C₃₁H₂₉N₄O₇Cl

[0598] Requires: C, 61.54; H, 4.83; N, 9.26;

[0599] Found: C, 61.55; H, 5.09; N, 9.14%.

Example 3N-(3,4,5-Trimethoxyphenyl)-3-(1-t-Butyl-pyrazolo[3,4-c]-1,2-Dihydro-6-Chloroquinolin-2-on-1-yl)phenylacetamide

[0600]

[0601] To a solution ofN-(3,4,5-trimethoxyphenyl)3-(1-(tert-butyl)-3-(2-chloro-5-fluoro-phenyl)pyrazol-4-oyl)aminophenylacetamide (595 mg, 1 mmol) in DMF (10 mL) powdered potassium carbonate(500 mg) was added and stirred at 100° C. overnight. The reactionmixture was diluted with ethyl acetate (200 ml) and washed with water(3×100 ml), brine, dried over sodium sulfate, evaporated andchromatographed (silica column, 50% ethyl acetate in chloroform). Yield500 mg, 86%.

[0602] NMR (CDCl₃): δ 1.70 (s, 9H, t-Bu), 3.78 (s, 2H, CH₂), 3.80 (s,6H, 2 OMe), 3.81 (s, 3H, OMe), 6.59 (d, 1H, Ar), 6.91 (s, 2H, (OMe)₃Ar),7.09 (t, 1H, Ar), 7.19 (d, 2H, Ar), 7.28 (t, 1H, Ar), 7.40 (d, 1H, Ar),7.52 (t, 1H, Ar), 8.25 (s, 1H, pyr).

[0603] MS(ES): 575 (M), 576 (M+1), 593 (M+18),

[0604] MS(ES): 634 (M+59).

Example 4N-(3,4,5-Trimethoxyphenyl)-3-(pyrazolo[3,4-c]-1,2-Dihydro-6-Chloroquinolin-2-on-1-yl)phenylacetamide

[0605]

[0606] A solution ofN-(3,4,5-trimethoxyphenyl)-3-(1-t-Butyl-pyrazolo[3,4-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide(46 mg, 0.8 mmol) in TFA reagent (2.5% anisole, 2.5% triisopropylsilane, 2.5% water and 92.5% TEA, 2 mL) was refluxed overnight. Thereaction mixture was evaporated and azeotroped with toluene, and theresidue was chromatographed (silica column, ethyl acetate). Yield 40 mg,100%.

[0607] NMR (CDCl₃): δ3.70 (s, 2H, CH₂), 3.80 (s, 3H, OMe), 3.82 (s, 6H,2 OMe), 6.79 (d, 1H, Ar), 6.91 (s, 2H, (OMe)₃Ar), 7.20-7.30 (m, 2H, Ar),7.39 (d, 1H, Ar), 7.49 (d, 1H, Ar), 7.60 (t, 1H, Ar), 7.89 (s, 1H, Ar),8.29 (s, 1H, pyr). DMSOd-6 (80° C.): δ 3.60 (s, 3H, OMe), 3.72 (s, 6H, 2OMe), 3.72, 2H, CH₂), 6.56 (d, 1H, Ar, adjacent to chloro), 7.25 (d, 1H,Ar), 7.28 (s, 1H, Ar), 7.37 (t, 1H, Ar), 7.42 (d, 1H, Ar), 7.52 (d, 1H,Ar), 7.59 (t, 1H, Ar), 8.41 (s, 1H, pyr), 10.16 (s, 1H, NH), 14.10 (brs, 1H, NH)

[0608]¹³C NMR (DMSOd-6, 80° C.): δ 42.8, 55.6, 60.0, 96.7, 115.8, 123.7,127.7, 129.3, 129.7, 130.0, 130.0, 133.5, 134.6, 135.2, 137.9, 138.0,141.6, 152.7, 168.1, 168.5,

[0609] MS(ES): 519 (M), 520 (M+1), 537 (M+18)

[0610] MS(ES): 517 (M−2), 518 (M−1), 519 (M).

Example 5N-Methyl-N-3,4,5-trimethoxyphenyl-3-(1-t-butyl-pyrazolo[3,4-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide

[0611]

[0612] To a suspension of NaH (30 mg, 0.6 mmol, 60% oil dispersion) inDMF (5 mL) a solution ofN-(3,4,5-trimethoxyphenyl)₃-(1-t-butyl-pyrazolo[3,4-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide(180 mg, 0.31 mmol) in DMF (10 mL) was added and heated at 100° C. for30 min and cooled to room temperature. Then methyl iodide (84 mg, 0.6mmol) was added and the reaction was stirred overnight. The reactionmixture was diluted with ethylacetate (100 mL), and quenched with water.The ethyl acetate solution was washed with water, brine, dried oversodium sulfate, filtered, evaporated and the residue was chromatographed(silica column, ethyl acetate). Yield 50 mg, 27%.

[0613] NMR (CDCl₃): δ 1.75 (s, 9H, t-But), 3.22 (s, 3H, N-Me), 3.70 (s,6H, 2 OMe). 3.80 (s, 5H, OMe and CH₂), 6.30 (s, 2H, (OMe)₃Ar), 6.55 (d,1H, Ar), 7.09 (s, 1H, Ar), 7.10 (s, 1H, Ar), 7.11 (s, 1H, Ar), 7.18 (d,1H, Ar), 7.25 (d, 1H, Ar), 7.22 (1H, Ar), 7.42 (t, 1H, Ar), 8.40 (s, 1H,Pyr).

[0614] MS(ES): 589 (M), 590 (M+1), 607 (M+18).

Example 6N-Methyl-N-3,4,5-trimethoxyphenyl-3-(pyrazolo[3,4-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide

[0615]

[0616] A solution ofN-methyl-N-3,4,5-trimethoxyphenyl-3-(1-t-butyl-pyrazolo[3,4-c]-1,2-dihydrochloroquinolin-2-on-1-yl)phenylacetamidein TFA reagent (2.5% anisole, 2.5% triisopropyl silane, 2.5% water and92.5% TFA, 3 mL) was refluxed for 4 hours. The reaction mixture wasevaporated and the residue was azeotroped with toluene and trituratedwith hexane to yield a white solid. The solid was dissolved inchloroform and filtered through a cation exchange column (6 cc, VarianBond Elute, SCX) and eluted with methanol. Methanolic fractionscontaining the product were evaporated. Yield 35 mg, 83%.

[0617] NMR (CD₃OD): δ 3.22 (s, 3H, N-Me), 3.64 (s, 2H), 3.70 (s, 6H, 2OMe), 3.71 (s, 3H, OMe), 6.25 (s, 2H, (OMe)₃Ar), 6.62 (d, 1H, Ar), 7.08(d, 1H, Ar), 7.06 (s, 1H, Ar), 7.14 (d, 1H, Ar), 7.21 (d, 1H, Ar),7.31-7.36 (m, 1H, Ar), 7.49 (t, 1H, Ar), 8.65 (s, 1H, pyr), 7.70-7.75(m, 1H, Ar), 11.60 (br s, pyr-NH).

[0618] MS(ES)+: 533 (M), 551 (M+18).

Example 7N-Methyl-N-3,4,5-trimethoxyphenyl-3-(1-methyl-pyrazolo[3,4-c]-1,24-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide

[0619]

[0620] To a suspension of sodium hydride (0.15 mmol, 6 mg, 60%dispersion) in DMF (5 mL), a solution ofN-methyl-N-3,4,5-trimethoxyphenyl-3-(pyrazolo[3,4-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide(20 mg, 0.037 mmol) was added and stirred at room temperature for 4hours, methyl iodide (1 mL, 3 mmol) was added, and the reaction wasstirred for 3 hours. The reaction mixture was diluted with ethyl acetateand (100 mL) washed with water (2×50 mL), dilute HCl (2×50 mL), brine(2×50 mL), dried over sodium sulfate, filtered, evaporated andchromatographed (purified by silica TLC, 20×20 cm, 250 m thickness, 30%ethyl acetate in chloroform). Yield 3.5 mg, 17%.

[0621] NMR (CDCl₃): δ 3.25 (s, 3H, N-Me), 3.59 (s, 2H, CH₂), 3.73 (s,6H, 2 OMe), 3.83 (s, 3H, OMe), 4.22 (s, 3H, pyr-N-Me), 6.33 (s, 2H,(OMe)₃Ar), 6.59 (d, 1H, Ar), 7.10-7.17 (m, 2H, Ar), 7.21-7.31 (m, 3H,Ar), 7.46 (t, 1H, Ar), 8.23 (s, 1H, pyr).

[0622] MS(ES)+: 547 (M), 548 (M+1), 549 (M+2).

Example 8N-3,4,5-Trimethoxyphenyl-3-(1-methyl-pyrazolo[3,4-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide

[0623]

[0624] To a solution ofN-3,4,5-trimethoxyphenyl-3-(pyrazolo[3,4-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl)phenylacetamide(100 mg, 0.19 mmol) in DMF (10 mL), cesium carbonate (100 mg, 0.3 mmol),was added and stirred for 3 h. At the end of three hours, methyl iodide(42 mg, 0.3 mmol) was added and reaction was stirred overnight. Thereaction mixture was diluted with ethyl acetate and washed with water,brine, dried over sodium sulfate, filtered and evaporated. The residuewas chromatographed (HPLC using Technikrom, 10 micron, silica column50.8×250 mm, 5% methanol in 95% chloroform to 100% methanol, flow rate75 mL/min). Yield 9 mg (9%) following unoptimized prep HPLC.

[0625] NMR (CD₃OD): δ 3.64 (s, 2H), 3.70 (s, 6H, 2 OMe), 3.71 (s, 3H,OMe), 4.21 (s, 3H, N-me), 6.25 (s, 2H, (OMe)₃Ar), 6.62 (d, 1H, Ar), 6.95(s, 1H, Ar), 7.08 (d, 1H, Ar), 7.14 (d, 1H, Ar), 7.18 (d, 1H, Ar),7.20-7.30 (m, 1H, Ar), 7.38 (t, 1H, Ar), 7.50-7.65 (m, 1H, Ar), 8.50 (s,1H, pyr).

[0626] MS(ES): 533 (M+1).

Example 9N-3,4,5-trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0627]

[0628] ToN-(3,4,5-trimethoxyphenyl)3-(3-methyl-5-(2-chloro-5-fluoro-phenyl)isoxazol-4-oyl)aminophenylacetamide (20 mg, 0.036 mmol) in DMF (1 ml) was added finely powderedK₂CO₃ (200 mg, 1.4 mmol). The suspension was rapidly stirred at roomtemperature. Workup involved dilution with EtOAc and 1 N HCl, transferto a separatory funnel, and extraction of the aqueous 3× (EtOAc). Thecombined organics were washed with saturated bicarbonate, brine, thendried. Filtration and concentration afforded the crude tricycle (16 mg).Purification (Bond-Elut silica cartridge, 0.5 g, 2:1 EtOAc/hexane)provided a pure sample (7.5 mg) of final product.

[0629] NMR (CDCl₃) δ 7.2, 7.3, 7.47, 7.6 (4 m, 6H), 6.8 (s, 2H), 6.65(dd, 1H), 3.76, 3.78 (2s+m, 11H), 2.6 (s, 3H).

[0630] MS (+ES) in/z 533.8 (M+H)⁺, 550.8 (M+NH₃)⁺.

Example 10N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydroquinolin-2-on-1-yl]phenylacetamide

[0631]

[0632] A solution from preparation 17 (0.27 g, 0.50 mmol), DMF (5.0 ml),and NaOH in MeOH (2.5 ml, 2N) was heated to 60° C. for 2 h then cooledto room temperature. EtOAc and H₂O were added and the layers separated.The organic fraction was washed (H₂O and brine), dried (MgSO₄),filtered, and concentrated. Column chromatography (silica gel,acetone/CH₂Cl₂) gave the title compound (0.19 g, 73%).

[0633]¹H NMR (400 MHz, CDCl₃) δ 8.22 (dd, 1H, J=1.4, 7.8 Hz), 7.61 (t,1H, J=7.8 Hz), 7.35 (m, 2H), 7.27 (m, 2H), 6.82 (s, 2H), 6.64 (d, 1H,J=8.5 Hz), 3.82 (s, 3H), 3.79 (s, 3H), 3.78 (s, 3H), 3.76 (s, 2H), 2.86(s, 3H) ppm.

[0634] Anal. calc. for C₂₈H₂₅N₃O₆; C, 67.33%; H, 5.04%; H, 8.41%; foundC, 67.25%; H, 5.27%; N, 8.30%.

[0635] MS(FAB) (m/z) 500.3 [M+1].

Example 11N-3,4,5-Trimethoxyphenyl-3-[3-phenyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0636]

[0637] A mixture of the product from preparation 22 (0.160 g, 0.27mmol), DMF (5 ml), and NaOH (2.5 ml, 2N in methanol) were reactedovernight. Dilution with ethyl acetate, washing (H₂O then brine), drying(MgSO₄), filtration and concentration gave the crude product.Purification by column chromatography (silica gel, hexanes/ethyl acetategradient) afforded the title compound (0.102 g, 66%).

[0638]¹H NMR (400 MHz, CDCl₃) δ 8.46 (dd, 1H, J=1.0, 8.3 Hz), 7.60 (t,1H, J=7.8 Hz), 7.51-7.42 (m, 5H), 7.34-7.21 (m, 5H), 6.74 (s, 2H), 6.55(dd, 1H, J=1.0, 8.3 Hz), 3.74 (s, 6H), 3.72 (s, 5H) ppm.

[0639] MS(FAB) (m/z) 595.4 [M+1].

Examples 12 and 13N-3,4,5-trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c-1,2-dihydro6-fluoroquinolin-2-on-1-yl]phenylacetamide &N-3,4,5-trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-methoxyquinolin-2-on-1-yl]phenylacetamide

[0640]

[0641] In a manner similar to that in example 10, the product frompreparation 27 (0.109 g, 0.20 mmol), 2N NaOH in MeOH (2 ml), and DMF (4ml) provided 8c1 (35 mg, 34%) and 8c2 (10 mg, 9%) after radialchromatography (silica gel, CH₂Cl₂/acetone gradient).

[0642]N-3,4,5-trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-fluoroquinolin-2-on-1-yl]phenylacetamide:

[0643]¹H NMR (400 MHz, CDCl₃) δ 7.60 (t, 1H, J=7.8 Hz), 7.48 (d, 1H,J=7.8 Hz), 7.29-7.20 (m, 4H), 7.00 (t, 1H, J=8.8 Hz), 6.78 (s, 2H), 6.41(d, 1H, J=8.8 Hz), 3.79 (s, 6H), 3.75 (s, 5H), 2.84 (s, 3H) ppm.

[0644] MS(FAB) Calc. 518.1727, found 518.1727.

[0645]N-3,4,5-trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-methoxyquinolin-2-on-1-yl]phenylacetamide:

[0646]¹H NMR (400 MHz, CDCl₃) δ 7.60 (t, 1H, J=7.8 Hz), 7.47 (m, 1H),7.27-7.15 (m, 5H), 6.78 (s, 2H), 6.21 (d, 1H, J=8.3 Hz), 4.06 (s, 3H),3.80 (s, 6H), 3.77 (s, 2H), 3.75 (s, 3H), 2.83 (s, 3H) ppm.

[0647] MS(FAB) Calc. 530.1927, found 530.1922.

Example 14N-3,4,5-Trimethoxyphenyl-3-[3-hexyl-isoxazolo[4,5-c]-1,2-dihydro-4-chloroquinolin-2on-1-yl]phenylacetamide

[0648]

[0649] Treatment of the product from preparation 30 (0.175 g, 0.28 mmol)in DMF (6 ml) was treated with 2N NaOH in MeOH (3 ml) for 18 h. Dilutionwith EtOAc followed by washing (H₂O then brine), drying (MgSO₄),filtration, and concentration gave the crude product that was purifiedby column chromatography (silica gel, CH₂Cl₂/acetone gradient) to givethe title compound (0.102 g, 60%).

[0650]¹H NMR (400 MHz, CDCl₃) 8 (7.60 (t, 1H, J=7.8 Hz), 7.48 (d, 1H,J=7.8 Hz), 7.30 (d 1H, J=6.8 Hz), 7.22-7.16 (m, 4H), 6.77 (s, 2H), 6.55(d, 1H, J=7.8 Hz), 3.79 (s, 6H), 3.76 (s, 2H), 3.75 (s, 3H), 3.24 (dt,2H, J=2.5, 7.3 Hz), 1.80 (quintet, 2H, J=7.3 Hz), 1.30-1.20 (m, 4H),0.82 (t, 3H, J=6.8 Hz) ppm.

[0651] MS(FAB) calc. 604.2214, found 604.2218.

Examples 15 and 16N-3,4,5-trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-4-fluoroquinolin-2-on-1-yl]phenylacetamide& N-3,4,5-trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro4-methoxyquinolin-2-on-1-yl]phenylacetamide

[0652]

[0653] In a fashion similar to that for example 10, compound frompreparation 35 (70 mg, 0.13 mmol), dimethylformamide (7 ml), and 2 NNaOH in methanol (3.5 ml) were reacted to give 8e1 (30 mg, 45%) and 8e2(6 mg, 9%) after column chromatography (silica gel, CH₂Cl₂/acetonegradient).

[0654]N-3,4,5-trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-4-fluoroquinolin-2on-1-yl]phenylacetamide:

[0655]¹H NMR (400 MHz, CDCl₃) δ 8.17 (dd, 1H, J=5.9, 8.3 Hz), 7.62 (t,1H, J=7.8 Hz), 7.50 (d, 1H, J=7.3 Hz), 7.25-7.20 (m, 2H), 7.08 (s, 1H),6.94 (dt, 1H, J=2.5, 7.8 Hz), 6.78 (s, 2H), 6.31 (dd, 1H, J=2.4, 10.7Hz), 3.80 (s, 6H), 3.77 (s, 2H), 3.75 (s, 3H), 2.84 (s, 3H) ppm.

[0656] MS(ES) (m/z) 517.9 [M+1].

[0657]N-3,4,5-trimethoxyphenyl-3-[3-methyl-isoxazolo[4,1-1,2-dihydro-4-methoxyquinolin-2-on-1-yl]phenylacetamide:

[0658]¹H NMR (400 MHz, CDCl₃) δ8.10 (d, 1H, J=8.8 Hz), 7.60 (t, 1H,J=7.8 Hz), 7.46 (d, 1H, J=7.8 Hz), 7.25-7.20 (m, 2H), 7.10 (s, 1H), 6.78(m, 3H), 6.08 (d, 1H, J=2.0 Hz), 3.79 (s, 6H), 3.76 (m, 2H), 3.75 (s,3H), 3.65 (s, 3H), 2.80 (s, 3H) ppm.

[0659] MS(ES) (m/z) 529.9 [M+1].

Example 17N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-4-bromoquinolin-2-on-1-yl]phenylacetamide

[0660]

[0661] In a fashion similar to that described for example 10, theproduct from preparation 40 (50 mg, 0.084 mmol), 2N NaOH in methanol(2.5 ml), and dimethylformamide (5.0 ml) were reacted to give (32 mg,67%) after column chromatography (silica gel, hexanes/ethyl acetategradient).

[0662]¹H NMR (400 MHz, CDCl₃) δ 8.04 (d, 1H, J=8.8 Hz), 7.61 (t, 1H,J=7.8 Hz), 7.50 (d, 1H, J=7.3 Hz), 7.35 (dd, 1H, J=2.0, 8.3 Hz), 7.24(m, 2H), 7.12 (s, 1H), 6.79 (s, 2H), 6.76 (d, 1H, J=2.0 Hz), 3.79 (s,6H), 3.78 (s, 2H), 3.75 (s, 3H), 2.82 (s, 3H) ppm.

[0663] MS(ES) (m/z) 578.0 [M+1], 580.0 [M+3].

Example 18N-3,4,5-Trimethoxyphenyl-3-[3-methyl(tetahydropyran-2-yl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0664]

[0665] To a solution from preparation 42 (1.05 g, 1.60 mmol) in DMF (30ml) was added powdered K₂CO₃ (2 g) at room temperature. After stirring18 h, ethyl acetate was added and the mixture was washed (H₂O thenbrine), dried (MgSO₄), filtered, and concentrated. Chromatography(silica gel, hexanes/dichloromethane/ethyl acetate gradient) gave thetitle compound (0.84 g, 83%).

[0666]¹H NMR (400 MHz, CDCl₃) δ 7.60 (t, 1H, J=7.8 Hz), 7.48 (d, 1H,J=7.8 Hz), 7.32 (dd, 1H, J=1.0, 7.8 Hz), 7.30-7.20 (m, 4H), 6.78 (s,2H), 6.56 (1H, dd, J=1.0, 8.8 Hz), 5.21 (dd, 1H, J=2.9, 14.2 Hz), 5.10(dd, 1H, J=2.9, 14.2 Hz), 4.85 (t, 1H, J=3.4 Hz), 3.86 (ddd, 1H, J=2.9,9.3, 12.2 Hz), 3.79 (s, 6H), 3.75 (s, 5H), 3.50 (ddd, 1H J=3.9, 5.4,10.7 Hz), 1.80-1.40 (m, 6H) ppm.

[0667] MS(ES) (m/z) 632.2 [M−1].

Example 19N-3,4,5-Trimethoxyphenyl-3-[3-hydroxymethyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0668]

[0669] To a solution from example 18 (0.830 g, 1.3 mmol) in methanol (50ml) and dichloromethane (25 ml) was added p-toluenesulfonic acid hydrate(0.070 g). After stirring 2 h, ethyl acetate was added and the mixturewashed with H₂O, saturated sodium bicarbonate, brine, dried (MgSO₄),filtered, and concentrated. Column chromatography (silica gel,dichloromethane/acetone) gave the title compound (0.702 g, 98%).

[0670]¹H NMR (400 MHz, CDCl₃) δ 7.63 (t, 1H, J=7.8 Hz), 7.53 (d, 1H,J=7.8 Hz), 7.37 (dd, 1H, J=1.0, 7.8 Hz), 7.28 (d, 1H, 8.3 Hz), 7.27-7.17(m, 3H), 6.75 (s, 2H), 6.61 (d, 1H, J=7.8 Hz), 5.43 (t, 1H, J=6.8 Hz),5.07 (d, 2H, J=6.8 Hz), 3.78 (s, 6H), 3.77 (s, 2H), 3.75 (s, 3H) ppm.

[0671] MS(EI) (m/z) 548.2 [M−1].

Example 20N-3,4,5-Trimethoxyphenyl-3-[3-azidomethyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0672]

[0673] To a solution of example 19 (106 mg, 0.193 mmol) and triethylamine (0.161 ml, 1.16 mmol) in dichloromethane (N₂, 0° C.) was addedmethane sulfonyl chloride (0.075 ml, 0.77 mmol). The reaction was warmedto room temperature after 10 minutes and diluted with dichloromethaneand water. The separated organic fraction was washed (brine), dried(MgSO₄), filtered, and concentrated to give the crude mesylate which wasstripped down with toluene (twice). The mesylate and sodium azide (75mg, 1.2 mmol) were dissolved in DMP (3 ml) at room temperature. After 5minutes ethyl acetate was added and the mixture washed with saturatedNaHCO₃, H₂O, brine, filtered, and concentrated. Column chromatography(silica gel, 10% acetone in dichloromethane) gave the title compound (83mg, 72%).

[0674]¹H NMR (400 MHz, CDCl₃) δ 7.60 (t, 1H, J=7.8 Hz), 7.50 (d, 1H, 7.8Hz), 7.34 (m, 2H), 7.26 (d, 1H, J=8.3 Hz), 7.21 (m, 2H), 6.78 (s, 2H),6.59 (d, 1H, J=8.2 Hz), 4.89 (s, 2H), 3.78 (s, 6H), 3.75 (s, 5H) ppm.

[0675] MS(ES) (m/z) 575.2 [M+1].

Example 21N-3,4,5-Trimethoxyphenyl-3-(3-1-butoxycarbonylaminomethyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0676]

[0677] A mixture of example 20 (75 mg, 0.130 mmol), (BOC)₂O (34 mg, 0.16mmol), Lindlar's catalyst (25 mg), and ethyl acetate (2 ml) was stirredunder one atmosphere of hydrogen gas (balloon) for 6 h, filtered throughcelite, and concentrated. Column chromatography (silica gel,dichloromethane/acetone gradient) gave the title compound (61 mg, 73%).

[0678]¹H NMR (400 MHz, CDCl₃) δ 7.60 (t, 1H, J=7.3 Hz), 7.50 (d, 1H,J=7.8 Hz), 7.31 (d, 1H, J=7.8 Hz), 7.29 (m, 1H), 7.24-7.16 (m, 4H), 6.77(s, 2H), 6.56 (d, J=8.3 Hz), 5.90 (br s, 1H), 4.83 (m, 2H), 3.77 (s,6H), 3.75 (s, 2H), 3.74 (s, 3H), 1.38 (s, 9H) ppm.

[0679] MS(ES) (m/z) 647.5 [M−1].

Example 22N-3,4,5-Trimethoxyphenyl-3-[3-amidylmethyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0680]

[0681] To a solution of example 21 (5.0 mg, 0.0077 mmol) indichloromethane (0.5 ml) at room temperature was added trifluoroaceticacid (0.5 ml). After 5 minutes the volatiles were removed and theresidue was applied to a SCX cation exchange cartridge. Elution withdichloromethane/methanol followed by 2M ammonia in methanol afforded thetitle compound (2.4 mg, 57%).

[0682]¹H NMR (400 MHz, CDCl₃) δ7.60 (t, 1H, J=7.8 Hz), 7.51 (d, 1H,J=7.3 Hz), 7.36 (s, 1H), 7.32 (d, 1H, J=7.8 Hz), 7.23-7.18 (m, 3H), 6.78(s, 2H), 6.57 (d, 1H, J=8.3 Hz), 4.34 (s, 2H), 3.79 (s, 6H), 3.77 (s,2H), 3.75 (s, 3H) ppm. MS(ES) (m/z) 549.2 [M+1].

Example 23N-3,4,5-Trimethoxyphenyl-3-[3-mesylmethyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0683]

[0684] A solution of the product from preparation 55 (47 mg, 0.075mmol), potassium phthalimide (50 mg, 0.27 mmol), and DMF (2 ml) wereallowed to react (17 h) at room temperature. The mixture was dilutedwith EtOAc then washed (H₂O, brine) and dried (MgSO₄). Columnchromatography (silica gel, CH₂Cl₂/acetone gradient) gave the titlecompound (27 mg, 53%).

[0685]¹H NMR (400 MHz, CDCl₃) δ 7.82 (m, 2H), 7.70 (m, 2H), 7.56 (t, 1H,J=7.8 Hz), 7.48 (d, 1H, J=7.8 Hz), 7.36 (s, 1H), 7.26 (d, 1H, J=6.8 Hz),7.24-7.18 (m, 3H), 6.76 (s, 2H), 6.55 (d, 1H, J=8.3 Hz), 5.41 (s, 2H),3.76 (s, 6H), 3.72 (s, 5H) ppm.

[0686] MS(ES) (m/z) 679.2 [M+1].

Example 24N-3,4,5-Trimethoxyphenyl-3-[3-methylthiomethyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0687]

[0688] A solution of the product from preparation 55 (50 mg, 0.080mmol), sodium thiomethoxide (20 mg, 0.29 mmol), and DMF (2 ml) wereallowed to react (17 h) at room temperature. The mixture was dilutedwith EtOAc then washed (H₂O, brine) and dried (MgSO₄). Columnchromatography (silica gel, CH₂Cl₂]acetone gradient) gave the titlecompound (41 mg, 82%).

[0689]¹H NMR (400 MHz, CDCl₃) δ 7.70-7.50 (m, 3H), 7.36 (t, 1H, J=7.8Hz), 7.28-7.21 (m, 3H), 6.83 (s, 2H), 4.23 (s, 2H), 3.81 (s, 6H), 3.79(s, 5H), 2.25 (s, 3H) ppm.

[0690] MS(ES) (m/z) 578 [M−1], 532 [M−SMe].

Example 25N-3,4,5-Trimethoxyphenyl-3-[3-thiazol-3-ylmethyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0691]

[0692] A solution of preparation 55 (26 mg, 0.041 mmol), thiazolidine(0.025 ml, 0.28 mmol), and DMF (2 ml) were allowed to react (17 h) atroom temperature. The mixture was diluted with EtOAc then washed (H₂O,brine) and dried (MgSO₄). Column chromatography (silica gel,CH₂Cl₂/acetone gradient) gave the title compound etamide (8.6 mg, 34%).

[0693]¹H NMR (400 MHz, CDCl₃) δ 7.58 (t, 1H, J=7.8 Hz), 7.50 (d, 1H,=7.8 Hz), 7.32 (dd, 1H, J=1.0, 7.8 Hz), 7.27-7.21 (m, 4H), 6.76 (s, 2H),6.56 (dd, 1H, J=1.0, 8.8 Hz), 4.19 (s, 2H), 4.16 (s, 2H), 3.78 (s, 6H),3.75 (s, 5H), 3.15 (t, 2H, J=6.8 Hz), 2.91 (t, 2H, J=6.8 Hz) ppm.

[0694] MS(ES) (m/z) 621.1 [M+1].

Example 26N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-iodoquinolin-2-on-1-yl]phenylacetamide

[0695]

[0696] To a solution of preparation 48 (20(mg, 0.32 mmol) in DMF (5 ml)was added powdered K₂CO₃ (88 mg, 0.64 mmol, 2 eq.) and stirred for 48 h.Partitioned between ethyl acetate and 0.1N HCl. Washed organic layerwith H₂O and brine, dried (MgSO₄), filtered and concentrated. The crudematerial was purified by flash chromatography using 1-15% acetone indichloromethane to give the title compound (170 g, 88%).

[0697] MS(ES) (m/z) 626.1 [M+1].

[0698] IR (KBr) 1679.56, 1632.05, 1606.52, 1587.85, 1508.40, 1411.66,1132.34 cm⁻¹.

Example 27N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-(thien-2-yl)-quinolin-2-on-1-yl]phenylacetamide

[0699]

[0700] To a mixture of example 26 (100 mg, 0.16 mmol) andtetrakis(triphenylphosphine)palladium (37 mg, 0.03 mmol, 20% mol) in THF(5 ml) under N₂, was added thiophene-2-boronic acid (51 mg, 0.4 mmol,2.5 eq.) dissolved in minimal ethanol and 2M Na₂CO₃ (0.16 ml, 2 eq.),and refluxed for 17 h. The mixture was cooled to room temperature,filtered, concentrated, and partitioned between ethyl acetate and H₂O.The organic layer was washed with brine, dried (MgSO₄), filtered andconcentrated. The crude material was filtered through a silica gel plugusing 1-10% acetone in dichloromethane to elute product then furtherpurified by HPLC (hexanes/ethyl acetate gradient) to give the titlecompound (29 mg, 31%).

[0701] MS(ES) (m/z) 582.2 [m+1].

[0702] IR (KBr) 1673.21, 1632.48, 1607.11, 1590.64, 1508.38, 1412.20,1132.29 cm⁻¹.

Example 28N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-(naphth-1-yl)-quinolin-2-on-1-yl]phenylacetamide

[0703]

[0704] To a mixture of example 26 (100 mg, 0.16 mmol) andtetrakis(triphenylphosphine)palladium (37 mg, 0.03 mmol, 20% mol), inTHF (5 ml) under N₂ was added 1-napthylboronic acid (69 mg, 0.4 mmol,2.5 eq.) dissolved in minimal ethanol and 2M Na₂CO₃ (0.16 ml, 2 eq.),and refluxed for 17 h. The mixture was cooled to room temperature,filtered, concentrated, and partitioned between ethyl acetate and H₂O.The organic layer was washed with brine, dried (MgSO₄), filtered andconcentrated. The crude material was filtered through a silica gel plugusing 1-10% acetone in dichloromethane to elute then purified by HPLC(hexanes/ethyl acetate gradient) to give the title compound (15 mg,15%).

[0705] MS(ES) (m/z) 626.2 [M+1].

[0706] IR (KBr) 1671.79, 1633.18, 1606.98, 1590.77, 1508.30, 1412.31,1132.32 cm⁻¹.

Example 29N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-(4-methoxyphenyl)quinolin-2-on-1-yl]phenylacetamide

[0707]

[0708] To a mixture of example 26 (100 mg, 0.16 mmol),tetrakis(triphenylphosphine)palladium (28 mg, 0.024 mmol, 15% mol), inTHF (5 ml) under N₂ was added 4-methoxyphenylboronic acid (49 mg, 0.32mmol, 2 eq.) dissolved in minimal ethanol, 2M Na₂CO₃ (0.16 ml, 2 eq.),and reaction was refluxed for 17 h. It was then cooled to roomtemperature, filtered, concentrated, and partitioned between ethylacetate and H₂O. The organic phase was washed with brine, dried (MgSO₄),filtered and concentrated. The crude material was run through a silicagel plug using 1-10% acetone in dichloromethane to elute then purifiedby HPLC (hexanes/ethyl acetate gradient) (2×) to give the title compound(24 mg, 25%).

[0709] MS(ES) (m/z) 606.3 [M+1].

[0710] IR (KBr) 1672.10, 1632.51, 1608.78, 1592.28, 1508.30, 1245.67,1245.67, 1132.33 cm⁻¹.

Example 30N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-(4-chlorophenyl)-quinolin-2-on-1-yl]phenylacetamide

[0711]

[0712] To a mixture of example 26 (100 mg, 0.16 mmol),tetrakis-(triphenylphosphine) palladium (28 mg, 0.024 mmol, 15% mol), inTHF (5 ml) under N₂ was added 4 chlorophenylboronic acid (50 mg, 0.32mmol, 2 eq.) dissolved in minimal ethanol, 2M Na₂CO₃ (0.16 ml, 2 eq.),and the reaction was refluxed for 17 h. It was then cooled to roomtemperature, filtered, concentrated, and partitioned between ethylacetate and H₂O. The organic layer was washed with brine, dried (MgSO₄),filtered and concentrated. The crude material was run through a silicagel plug using 1-10% acetone in dichloromethane to elute then purifiedby HPLC (hexanes/ethyl acetate gradient) (2×) to give the title compound(28 mg, 29%).

[0713] MS(ES) (m/z) 610.3 [M+1].

[0714] IR(KBr) 1675.84, 1632.45, 1606.41, 1591.95, 1508.06, 1477.21,1411.64, 1132.01 cm⁻¹.

Example 31N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-(ethylenyl)-quinolin-2-on-1-yl]phenylacetamide

[0715]

[0716] To a mixture of example 26 (100 mg, 0.16 mmol) and PdCl₂(PPh₃)₄(17 mg, 0.024 mmol, 15% mmol) in THF (3 ml) was added tributylvinyl tin(0.05 ml, 0.18 mmol, 1.1 eq.), and the reaction was refluxed for 24 h.After cooling to room temperature, it was partitioned between ethylacetate and H₂O and the organic layer was washed with brine, dried(MgSO₄), and concentrated. The crude material was run through a silicagel plug using ethyl acetate, then purified by HPLC (hexanes/ethylacetate gradient) to give the title compound (10 mg, 12%).

[0717] MS(ES) (m/z) 526.3 [M+1].

[0718] IR (KBr) 3286.85, 2933.78, 1668.15, 1606.59, 1543.09, 1507.12,1410.36, 1329.45, 1231.00, 1128.87, 1008.70 cm⁻¹.

Example 32N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-benzyl-quinolin-2-on-1-yl]phenylacetamide

[0719]

[0720] To a mixture of 0.5M benzyl-9-BBN (1 ml) in DMF (2 ml) under N₂was added PdCl₂ (26 mg, 0.032 mmol, 20% mmol), the product from example26 (100 mg, 0.16 mmol), and powdered K₂CO₃ (44 mg, 0.32 mmol, 2 eq.). Itwas then heated at 50° C. for 24 h. then cooled to room temperature. IceH₂O was added and extracted with ethyl acetate (2×). The combinedorganic layers were washed with H₂O brine, dried (MgSO₄), filtered andconcentrated. The crude material was run through a silica gel plug using1-10% acetone in dichloromethane then purified by HPLC (hexanes/ethylacetate gradient) to give the tide compound (4 mg, 4%).

[0721]¹H NMR (400 MHz, CDCl₃) δ 7.63 (t, 1H), 7.50 (d, 1H), 7.30-7.21(m, 8H), 6.97 (d, 1H), 6.82 (s, 2H), 6.53 (d, 1H), 4.72 (s, 2H), 3.82(s, 6H), 3.79 (s, 3H), 2.86 (t, 3H) ppm.

[0722] MS(ES) (m/z) 590.4 [M+1].

Example 33N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-methoxycarbonyl-quinolin-2-on-1-yl]phenylacetamide

[0723]

[0724] A mixture of example 26 (200 mg, 032 mmol), Pd(OAc)₂ (22.5 mg),methanol (7 ml), acetonitrile (30 ml), and triethylamine (0.16 ml) washeated to 60° C. for 24 h with 60 psi Carbon monoxide. After celitefiltration and concentration, the crude material was run through asilica gel plug using 1-10% acetone in dichloromethane, then purified byHPLC (hexanes/ethyl acetate gradient) to give the title compound (44 mg,25%).

[0725] MS(ES) (m/z) 558.1 [M+1].

[0726] IR (KBr) 1735.99, 1685.23, 1632.98, 1605.89, 1507.07, 1486.61,1450.39, 1410.73, 1313.11, 1290.07, 1231.18, 1127.96 cm⁻¹.

Example 34N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-6-carboxy-quinolin-2-on-1-yl]phenylacetamide

[0727]

[0728] A solution of example 33 (20 mg, 0.04 mmol) in methanol (0.5 ml)was treated with 1N NaOH (0.07 ml, 2 eq.) and heated at 50° C. for 24 h.The reaction was cooled to room temperature and ice H₂O was added.Organic impurities were removed with hexanes, then the solution wasacidified to pH 2 and extracted with ethyl acetate (2×). The organiclayer was washed with H₂O, then brine, dried (MgSO₄), filtered, andconcentrated to give the title compound (18 mg, 92%).

[0729] MS(ES) (m/z) 544.3 [M+1].

[0730] IR (KBr) 3271.51, 2935.24, 1717.49, 1672.13, 1632.11, 1595.29,1536.64, 1506.49, 1451.73, 1412.95, 1332.17, 1225.33, 1129.35, 1007.60,813.62 cm⁻¹.

Example 35N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-1,2-dihydro-5-iodoquinolin-2-on-1-yl]phenylacetamide

[0731]

[0732] To a solution of preparation 54 (200 mg, 0.32 mmol) in DMF (5 ml)was added powdered K₂CO₃ (88 mg, 0.64 mmol, 2 eq.) and stirred for 17 h.The reaction was partitioned between ethyl acetate and 0.1N HCl, andwashed organic layer with H₂O and brine, dried (MgSO₄), filtered andconcentrated. The crude material was purified by flash chromatography(silica) using 1-20% acetone in dichloromethane to give the titlecompound (168 g, 87% o).

[0733] MS(ES) (m/z) 626.1 [M+1].

[0734] IR (KBr) 1678.85, 1630.32, 1603.94, 1508.59, 1484.62, 1132.20cm⁻¹.

Example 36N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-5-(4-trifluoromethylphenyl)quinolin-2-on-1-yl]phenylacetamide

[0735]

[0736] To a mixture of example 35 (100 mg, 0.16 mmol),tetrakis(triphenylphosphine)palladium (28 mg, 0.024 mmol, 15% mol), inTHF (5 ml) under N₂ was added 4-Trifluoromethylphenylboronic acid (50mg, 0.32 mmol, 2 eq.) dissolved in minimal ethanol, 2M Na₂CO₃ (0.16 ml,2 eq.), and refluxed for 24 h. Rxn was cooled to room temperature,filtered, concentrated, and partitioned between ethyl acetate and H₂O.The organic layer was washed with brine, dried (MgSO₄), filtered andconcentrated. The crude material was run through a silica gel plug using1-15% acetone in dichloromethane to elute then purified by HPLC(hexanes/ethyl acetate gradient) to the tide compound (46 mg, 45%).

[0737] MS(ES) (m/z) 644.3 [M+1].

[0738] IR (KBr) 1679.00, 1617.95, 1508.45, 1325.99, 1132.03 cm⁻¹.

Example 37N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-5-naphthyl-quinolin-2-on-1-yl]phenylacetamide

[0739]

[0740] To a mixture of example 35 (100 mg, 0.16 mmol),tetrakis(triphenylphosphine) palladium (28 mg, 0.024 mmol, 15% mol), inTHF (5 ml) under N₂ was added 1-napthylboronic acid (50 mg, 0.32 mmol, 2eq.) dissolved in minimal ethanol, 2M Na₂CO₃ (0.16 ml, 2 eq.), andrefluxed for 24 h. The reaction was cooled to room temperature,filtered, concentrated, and partitioned between ethyl acetate and H₂O.Washed organic layer with brine, dried (MgSO₄), filtered andconcentrated. The crude material was run through a silica gel plug using1-15% acetone in dichloromethane, then purified by HPLC (hexanes/ethylacetate gradient) to give the title compound (70 mg, 70%).

[0741] MS(ES) (m/z) 610.3 [M+1].

[0742] IR (KBr) 1672.33, 1634.42, 1619.23, 1607.85, 1508.41, 1498.70,1452.75, 1412.20, 1237.11, 1132.15 cm⁻¹.

Example 38N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-5-(thien-2-yl)-quinolin-2-on-1-yl]phenylacetamide

[0743]

[0744] To a mixture of example 35 (100 mg, 0.16 mmol),tetrakis(triphenylphosphine)palladium (28 mg, 0.024 mmol, 15% mol), inTHF (5 ml) under N₂ was added thiophene-2-boronic acid (41 mg, 0.32mmol, 2 eq.) dissolved in minimal ethanol, 2M Na₂CO₃ (0.16 ml, 2 eq.),and refluxed for 24 h. The reaction was cooled to room temperature,filtered, concentrated, and partitioned between ethyl acetate and H₂O.The mixture was washed with brine, dried (MgSO₄), filtered andconcentrated. The crude material was run through a silica gel plug using1-15% acetone in dichloromethane, then purified by HPLC (hexanes/ethylacetate gradient) to give the title compound (52 mg, 56%)].

[0745] MS(ES) (m/z) 582.2 [M+1].

[0746] IR (KBr) 1675.07, 1619.94, 1607.44, 1508.48, 1495.78, 1132.25cm⁻¹.

Example 39N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-5-(4-methoxyphenyl)quinolin-2-on-1-yl]phenylacetamide

[0747]

[0748] To a mixture of example 35 (100 mg, 0.16 mmol),tetrakis(triphenylphosphine)palladium (6 mg, 0.006 mmol, 3% mol), in THF(4 ml) under N₂ was added 4-methoxyphenylboronic acid (27 mg, 0.18 mmol,1.1 eq.) dissolved in minimal ethanol, 2M Na₂CO₃ (0.18 ml, 1.1 eq.), andrefluxed for 24 h. The reaction was cooled to room temperature,filtered, concentrated, and partitioned between ethyl acetate and H₂O.The organic layer was washed with brine, dried (MgSO₄), filtered andconcentrated. The crude material was run through a silica gel plug using1-15% acetone in dichloromethane, then purified by HPLC (hexanes/ethylacetate gradient) to give the title compound (33 mg, 34%).

[0749] MS(ES) (m/z) 606.3 [M+1].

[0750] IR (KBr) 1674.98, 1619.77, 1608.84, 1508.35, 1491.99, 1132.22cm⁻¹.

Example 40N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-5-(4-chlorophenyl)-quinolin-2-on-1-yl]phenylacetamide

[0751]

[0752] To a mixture of example 19 (100 mg, 0.16 mmol),tetrakis(triphenylphosphine)palladium (6 mg, 0.006 mmol, 3% mol), in THF(4 ml) under N₂ was added 4-chlorophenylboronic acid (28 mg, 0.18 mmol,1.1 eq.) dissolved in minimal ethanol, 2M Na₂CO₃ (0.18 ml, 1.1 eq.), andrefluxed for 24 h. The reaction was cooled to room temperature,filtered, concentrated, and partitioned between ethyl acetate and H₂O.The organic layer was washed with brine, dried (MgSO₄), filtered andconcentrated. The crude material was run through a silica gel plug using1-10% acetone in dichloromethane, then purified by HPLC (hexanes/ethylacetate gradient) to give the title compound (38 mg, 39%).

[0753] MS(ES) (m/z) 610.1 [M+1].

[0754] IR (KBr) 1675.47, 1619.83, 1508.75, 1485.81 cm⁻¹.

Example 41N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-5-(methoxycarbonyl)quinolin-2-on-1-yl]phenylacetamide

[0755]

[0756] A mixture of example 35 (100 mg, 016 mmol), Pd(OAc)₂ (18 mg),methanol (5 ml), acetonitrile (25 ml), and triethylamine (0.13 ml) washeated to 60° C. for 24 h under 60 psi Carbon monoxide. The crude wasfiltered through a celite plug and concentrated. The crude material waspurified by flash chromatography using 1-10% acetone in dichloromethaneto give the title compound (41 mg, 46%).

[0757] MS(ES) (m/z) 558.2 [M+1].

[0758] IR (KBr) 1721.23, 1684.85, 1608.69, 1619.50, 1508.33, 1412.22,1274.85, 1412.22, 1274.85, 1233.15, 1131.68 cm⁻¹.

Example 42N-3,4,5-Trimethoxyphenyl-3-[3-methyl-isoxazolo[4,5-c]-1,2-dihydro-5-carboxy-quinolin-2-on-1-yl]phenylacetamide

[0759]

[0760] A solution of example 41 (25 mg, 0.05 mmol) in methanol (0.5 ml)and THF (0.25 ml) was treated with 1N NaOH (0.17 ml, 2 eq.) and heatedat 50° C. for 2 h 30 min. The reaction was cooled to room temperature,diluted with ice H₂O, and organic impurities were extended with hexanes.The aqueous layer was acidified to pH 2, then extracted with ethylacetate (2×). The organic layer was washed with H₂O, brine, dried(MgSO₄), filtered, and concentrated to give the title compound (24 mg,100%).

[0761] MS(ES) (m/z) 542.3 [M−1].

[0762] IR (KBr) 3356.3, 2945.2, 1675.05, 1618.10, 1506.87, 1234.68,1128.89 cm⁻¹.

Example 43N-3,4,5-Trimethoxyphenyl-3-[3-methylthio-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0763]

[0764] To a solution of preparation 5 (17 mg, 0.03 mmol) in DMF (3 ml)under N₂ was added sodium methylthiolate (11 mg, 0.15 mmol, 5 eq.). Thesolution was stirred for 45 min., partitioned between ethyl acetate andH₂O. The organic layer was washed with brine, dried (MgSO₄), filteredand concentrated. The crude material was purified by flashchromatography using 1-10% acetone in dichloromethane to give the titlecompound (6 mg, 35%).

[0765] MS(FAB) (m/z) 561.1.

[0766] IR (KBr) 1679.98, 1656.59, 1600.17, 1578.25, 1507.22, 1460.51,1455.02, 1410.12, 1229.79, 1136.54.

Example 44N-3,4,5-Trimethoxyphenyl-3-[3-chloroisoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0767]

[0768] To a solution of preparation 5 (0.5 g, 0.88 mmol) in DM (50 ml)under N₂ was added dropwise 1.0M sodium trimethylsilanolate (0.9 ml,1.05 eq.) over 3.6 h. The mixture was partitioned between ethyl acetateand 0.1N HCl, and the organic layer was washed with H₂O, brine, dried(MgSO₄), filtered and concentrated. The crude material was purified byflash chromatography using 10-75% ethyl acetate in dichloromethane togive the tide compound (224 mg, 46%).

[0769] Anal. Calc. for C₂₇H₂₁Cl₂N₃O₆

[0770] Theoretical: C, 58.50; H, 3.82; N, 7.58;

[0771] Found. C, 58.84; H, 5.05; N, 7.87%.

[0772] IR (KBr) 1673.09, 1593.79, 1459.25, 1132.05 cm⁻¹.

Example 45N-3,4,5-Trimethoxyphenyl-3-[3-(diethylamidyl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0773]

[0774] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (5 ml)under N₂ was added diethylamine (28 ul, 0.27 mmol, 3 eq.). The solutionwas stirred at room temperature for 30 min, then partitioned betweenethyl acetate and 0.1N HCl. The organic layer was washed with H₂O,brine, dried (MgSO₄), filtered and concentrated. The crude material waspurified by flash chromatography using 1-10% acetone in dichloromethaneto give the title compound (32 mg, 60%).

[0775]¹H NMR (400 MHz, CDCl₃) δ 7.58 (t, 1H), 7.47 (d, 1H), 7.44 (s,1H), 7.23-7.19 (m, 3H), 7.13 (t, 1H), 6.79 (s, 2H), 6.40 (d, 2H), 3.89(q, 4H), 3.79 (s, 6H), 3.78 (s, 3H), 3.73 (s, 2H), 1.26 (t, 6H) ppm.

[0776] MS(ES) (m/z) 591.4 [M+1].

Example 46N-3,4,5-Trimethoxyphenyl-3-[3-(dipropylamidyl)isoxazolo[4,5-c]1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0777]

[0778] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (5 ml)under N₂ was added dipropylamine (37 ul, 0.27 mmol, 3 eq.). The solutionwas stirred at room temperature for 30 nm, then partitioned betweenethyl acetate and 0.1N HCl. The organic layer was washed with H₂O,brine, dried (MgSO₄), filtered and concentrated. The crude material waspurified by flash chromatography using 1-10% acetone in dichloromethaneto give the title compound (20 mg, 36%). !H NMR (400 MHz, CDCl₃) δ 7.59(t, 1H), 7.47 (d, 1H), 7.41 (s, 1H), 7.23-7.19 (m, 3H), 7.13 (t, 1H),6.79 (s, 2H), 6.40 (d, 2H), 3.85-3.79 (m, 4H), 3.80 (s, 6H), 3.78 (s,3H), 3.75 (s, 2H), 1.70-1.64 (m, 4H), 0.88 (t, 6H) ppm.

[0779] MS(ES) (m/z) 619.5 [M+1].

Example 47N-3,4,5-Trimethoxyphenyl-3-[3-(4-methoxyamino)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0780]

[0781] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (55 ml)under N₂ was added p-anisidine (55 mg, 0.45 mmol, 5 eq.). The solutionwas stiffed at room temperature for 1 h 45 min, then partitioned betweenethyl acetate and 0.1N HCl. The organic layer was washed with H₂O,brine, dried (MgSO₄), filtered and concentrated. The crude material waspurified by flash chromatography using 1-15% acetone in dichloromethaneto give the title compound (26 mg, 45%).

[0782]¹H NMR (400 MHz, CDCl₃) δ 8.61 (s, 1H), 7.59 (t, 1H), 7.56 (s,1H), 7.53 (d, 1H), 7.32 (d, 2H), 7.27 (s, 1H), 7.22-7.18 (m, 3H), 6.87(d, 2H), 6.83 (s, 2H), 6.58 (d, 2H), 3.79 (s, 9H), 3.78 (s, 3H), 3.74(s, 2H) ppm.

[0783] MS(ES) (m/z) 663.162 [M+Na].

Example 48N-3,4,5-Trimethoxyphenyl-3-[3-(thiazol-3-yl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0784]

[0785] To a solution of example 44 (2 mg, 0.04 mmol) in DMF (2 ml) underN₂ was added thiazolidine (18 ul, 0.24 mmol, 6 eq.). The solution wasstirred at room temperature for 1 h, then partitioned between ethylacetate and H₂O. The organic layer was washed with brine, dried (MgSO₄),filtered and concentrated. The crude material was purified by flashchromatography using 1-10% acetone in dichloromethane to give the titlecompound (16 mg, 67%).

[0786]¹H NMR (400 MHz, CDCl₃) δ 7.61 (t, 1H), 7.51 (d, 1H), 7.27-7.25(m, 1H), 7.22-7.16 (m, 3H), 6.78 (s, 2H), 6.47 (d, 1H), 5.09 (s, 1H),4.26 (t, 2H) 3.81 (s, 6H), 3.79 (s, 3H), 3.77 (s, 2H), 3.11 (t, 2H) ppm.

[0787] MS(FAB) (m/z) 607.2 [M+1].

Example 49N-3,4,5-Trimethoxyphenyl-3-[3-tenzylthio-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0788]

[0789] To a solution of example 44 (20 mg, 0.04 mmol) in DMF (2 ml)under N₂ was added benzyl mercaptan (20 ul, 0.2 mmol, 5 eq.). Thesolution was stirred at room temperature for 18 h, then at 70° C. for 6h, and partitioned between ethyl acetate and H₂O. The organic layer waswashed with brine, dried (MgSO₄), filtered and concentrated The crudematerial was purified by flash chromatography using 1-15% acetone indichloromethane to give the title compound (15 mg, 58%).

[0790] IR (KBr) 3310.21, 3264.90, 2926.45, 2831.95, 1699.87, 1674.74,1613.05, 1592.37, 1506.49, 1458.07, 1409.78, 1230.24, 1128.31 cm⁻¹.

[0791] MS(FAB) (m/z) 642.1479.

Example 50N-3,4,5-Trimethoxyphenyl-3-[3-phthalimidyl-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetarmide

[0792]

[0793] To a solution of example 44 (20 mg, 0.04 mmol) in DMF (2 ml)under N₂ was added potassium phthalimide (37 mg, 0.2 mmol, 5 eq.). Thereaction was stirred at room temperature for 1 h, then partitionedbetween ethyl acetate and H₂O. The organic layer was washed with brine,dried (MgSO₄), filtered and concentrated. The crude material waspurified by flash chromatography using 1-10% acetone in dichloromethaneto give the title compound (10 mg, 42%).

[0794]¹H NMR (400 MHz, CDCl₃) δ 7.97-7.95 (m, 2H), 7.83-7.81 (m, 2H),7.57 (t, 1H), 7.50 (d, 1H), 7.38 (d, 1H), 7.31-7.27 (m, 2H), 7.23-7.22(m, 1H), 6.72 (s, 2H), 6.62 (d, 1H), 3.76 (s, 3H), 3.72 (s, 6H), 3.71(s, 2H) ppm.

[0795] MS(FAB) (m/z) 665.1439.

Example 51N-3,4,5-Trimethoxyphenyl-3-[3-pyridin-2-ylthio-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0796]

[0797] To a solution of example 44 (20 mg, 0.04 mmol) in DMF (2 ml)under N₂ was added 2-mercaptopyridine (22 mg, 0.2 mmol, 5 eq.). Thereaction was stirred at room temperature for 1 h, then partitionedbetween ethyl acetate and H₂O. The organic layer was washed with brine,dried (MgSO₄), filtered and concentrated. The crude material waspurified by flash chromatography using 1-15% acetone in dichloromethaneto give the desired compound (19 mg, 84%).

[0798]¹H NMR (400 MHz, CDCl₃) δ 8.60-8.58 (m, 1H), 7.75 (t, 1H), 7.72(d, 1H), 7.68 (t, 1H), 7.60 (d, 1H), 7.51 (s, 1H), 7.33-7.31 (m, 2H),7.26-7.22 (m, 2H), 6.84 (s, 2H), 6.62 (d, 1H), 3.82 (s, 6H), 3.77 (s,3H), 3.75 (s, 2H) ppm.

[0799] MS(FAB) (m/z) 629.1242.

Example 52N-3,4,5-Trimethoxyphenyl-3-[3-phenylthio-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0800]

[0801] To a solution of example 44 (4 mg, 0.007 mmol) in DMF (0.5 ml)under N₂ was added sodium phenylthiolate (5 mg, 0.035 mmol, 5 eq.). Thereaction was stirred at room temperature for 18 h, then at 60° C. for 3h, and partitioned between ethyl acetate and H₂O. The organic layer waswashed with brine, dried (MgSO₄), filtered and concentrated. The crudematerial was purified by flash chromatography using 1-10% acetone indichloromethane to give the desired compound (2 mg, 47%).

[0802]¹H NMR (400 MHz, CDCl₃) δ 7.69-7.63 (m, 3H), 7.53-7.45 (m, 4H),7.33-7.23 (m, 4H), 6.85 (s, 2H), 6.62 (d, 1H), 3.86 (s, 6H), 3.82 (s,2H), 3.78 (s, 3H) ppm.

[0803] MS(FAB) (m/z) 628.1313.

Example 53N-3,4,5-Trimethoxyphenyl-3-[3-((2-methoxy-1-hydroxy-ethan-2-al)amidyl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0804]

[0805] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (5 ml)under N₂ was added DL-serine methyl ester hydrochloride (70 mg, 0.45mmol, 5 eq.), then diisopropylethylamine (50 ul, 0.27 mmol, 3 eq). Itwas stirred at room temperature for 2 h, and then partitioned betweenethyl acetate and 0.1N HCl. The organic layer was washed with H₂O,brine, dried (MgSO₄), filtered and concentrated. The crude material waspurified by flash chromatography using 1-30% acetone in dichloromethaneto give the title compound (33 mg, 58%).

[0806] MS(ES) (m/z) 637.2 [M+1].

[0807] IR (KBr) 3676.62, 3361.32, 3017.09, 1748.50, 1669.24, 1600.15,1507.87, 1464.59, 1452.69, 1412.75, 1286.17, 1267.41, 1131.66, 1148.36cm⁻¹.

Example 54N-3,4,5-Trimethoxyphenyl-3-[3-(4-fluoroanilino)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0808]

[0809] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (5 ml)under N₂ was added 4-fluoroaniline (26 ul, 0.27 mmol, 3 eq.) thendiisopropylethylamine (50 ul, 0.27 mmol, 3 eq). The reaction was stirredat room temperature for 7 h, and partitioned between ethyl acetate and0.1N HCl. The organic layer was washed with H₂O, brine, dried (MgSO₄),filtered and concentrated. The crude material was purified by flashchromatography using 1-15% acetone in dichloromethane to give the titlecompound (37 mg, 66%).

[0810]¹H N (400 MHz, CDCl₃) δ 7.63-7.60 (m, 2H), 7.52 (d, 1H), 7.34 (m,2H), 7.28-7.18 (m, 3H), 7.05 (t, 2H), 6.83 (s, 2H), 6.59 (d, 1H), 3.79(s, 6H), 3.78 (s, 3H), 3.73 (s, 2H) ppm. MS(FAB) (m/z) 629.1611.

Example 55N-3,4,5-Trimethoxyphenyl-3-[3-anilino-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0811]

[0812] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (5 ml)under N₂ was added aniline (25 ul, 0.27 mmol, 3 eq.), thendiisopropylethylamine (50 ul, 0.27 mmol, 3 eq). The reaction was stirredat room temperature for 19 h, and partitioned between ethyl acetate and0.1N HCl. The organic layer was washed with H₂O, brine, dried (MgSO₄),filtered and concentrated. The crude material was purified by flashchromatography using 1-15% acetone in dichloromethane to give thedesired compound the title compound (34 mg, 61%).

[0813]¹H NMR (400 MHz, CDCl₃) δ 7.63 (t, 1H), 7.53 (d, 1H), 7.44-36 (m,5H), 7.30-7.21 (m, 3H), 7.15 (t, 1H), 6.82 (s, 2H), 6.61 (d, 1H), 3.81(s, 6H), 3.79 (s, 3H), 3.77 (s, 2H) ppm.

[0814] MS(FAB) (m/z) 611.1703.

Example 56N-3,4,5-Trimethoxyphenyl-3-[3-(2-methoxyethylamidyl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0815]

[0816] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (5 ml)under N₂ was added 2-methoxyethylamine (24 ul, 0.27 mmol, 3 eq.) thendiisopropylethylamine (50 ul, 0.27 mmol, 3 eq). The mixture was stirredat room temperature for 10 min, then partitioned between ethyl acetateand 0.1N HCl. The organic layer was washed with H₂O, brine, dried(MgSO₄), filtered and concentrated. The crude material was purified byflash chromatography using 1-30% acetone in dichloromethane to give thedesired compound the title compound (31 mg, 59%).

[0817] MS(FAB) (m/z) 593.1809.

[0818] IR (KBr) 1668.82, 1639.51, 1600.75, 1508.25, 1452.72, 1131.93cm⁻¹.

Example 57N-3,4,5-Trimethoxyphenyl-3-[3-((2-methoxy-1-prop-2-yl-ethan-2-al)amidyl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0819]

[0820] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (5 ml)under N₂ was added D,L-valine methyl ester hydrochloride (76 mg, 0.45mmol, 5 eq.), then diisopropylethylamine (50 ul, 0.27 mmol, 3 eq). Themixture was stirred at room temperature for 2 h, then partitionedbetween ethyl acetate and 0.1N HCl. The organic layer was washed withH₂O, brine, dried (MgSO₄), filtered and concentrated. The crude materialwas purified by flash chromatography using 1-15% acetone indichloromethane to give the title compound (38 mg, 66%), as a mixture ofenantiomers.

[0821] MS(FAB) (m/z) 649.2059.

[0822] IR (KBr) 3010.50, 1744.23, 1667.18, 1638.41, 1599.86, 1508.11,1464.67, 1452.64, 1412.44, 1285.83, 1266.55, 1235.18, 1149.15, 1132.61cm⁻¹.

Example 58N-3,4,5-Trimethoxyphenyl-3-[3-(2-methoxycarbonylpyrrolidinyl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0823]

[0824] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (5 ml)under N₂ was added L-proline methyl ester hydrochloride (75 mg, 0.45mmol, 5 eq.), then diisopropylethylamine (75 ul, 0.41 mmol, 4.5 eq). Thereaction was stirred at room temperature for 1 h 15 min, thenpartitioned between ethyl acetate and 0.1N HCl. The organic layer waswashed with H₂O, brine, dried (MgSO₄), filtered and concentrated. Thecrude material was purified by flash chromatography using 1-20% acetonein dichloromethane to give the title compound (56 mg, 90%).

[0825] MS(FAB) (m/z) 647.1896.

[0826] IR (KBr) 1744.62, 1669.01, 1603.27, 1594.03, 1498.34, 1452.51,1412.85, 1287.30, 1235.18, 1132.45 cm⁻¹.

Example 59N-3,4,5-Trimethoxyphenyl-3-[3-((2-methoxy-1-benzyl-ethan-2-al)amidyl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0827]

[0828] To a solution of example 44 (50 mg, 0.09 mmol) in DMF (5 ml)under N₂ was added D,L-phenylalanine hydrochloride (97 mg, 0.45 mmol, 5eq.), then diisopropylethylamine (75 ul, 0.41 mmol, 4.5 eq). It wasstirred at room temperature for 24 h, then partitioned between ethylacetate and 0.1N HCl. The organic layer was washed with H₂O, brine,dried (MgSO₄), filtered and concentrated. The crude material waspurified by flash chromatography using 1-15% acetone in dichloromethaneto give the title compound (41 mg, 65%) as a mixture of enantiomers.

[0829] MS(FAB) (m/z) 697.2075.

[0830] IR (KBr) 1748.03, 1668.31, 1638.93, 1599.78, 1508.08, 1452.64,1412.32, 1284.48, 1267.21, 1235.18, 1132.20 cm⁻¹.

Example 60N-3,4,5-Trimethoxyphenyl-3-[3-((2-methoxy-1-methyl-ethan-2-al)amidyl)-isoxazolo[4,5-c3-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0831]

[0832] To a solution of example 44 (36 mg, 0.07 mmol) in DMF (3.5 ml)under N₂ was added (D,L) ananine methyl ester (49 mg, 0.35 mmol, 5 eq.),then triethylamine (30 ul, 0.21 mmol, 3 eq.). The solution was stirredat room temperature for 3 h, then partitioned between ethyl acetate andH₂O. The organic layer was washed with brine, dried (MgSO₄), filteredand concentrated. The crude material was purified by flashchromatography using 1-10% acetone in dichloromethane to give the titlecompound (16 mg, 40%) as a mixture of enantiomers.

[0833] Anal. Calc. for C₃₁H₂₉ClN₄O₈

[0834] Theoretical: C, 59.95; H, 4.71; N, 9.02;

[0835] Found C, 59.90; H, 5.08, 8.52%.

[0836] IR (KBr) 1746.09, 1671.16, 1638.64, 1598.09, 1506.60, 1449.88,1410.66, 1280.66, 1267.05, 1229.78, 1151.16, 1128.33, 789.32 cm⁻¹.

Example 61N-3,4,5-Trimethoxyphenyl-3-[3-((2-methoxy-ethan-2-al)amidyl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0837]

[0838] To a solution of example 44 (23 mg, 0.04 mmol) in DMF (2 m])under N₂ was added triethylamine (261, 0.2 mmol, 5 eq.), then glycinemethyl ester (15 mg, 0.12 mmol, 3 eq.). The solution was stirred at roomtemperature for 3 h, and partitioned between ethyl acetate and H₂O. Theorganic layer was washed with brine, dried (MgSO₄), filtered andconcentrated. The crude material was purified by flash chromatographyusing 1-10% acetone in dichloromethane to give the tide compound (16 mg,40%).

[0839] MS(FAB) (m/z) 607.1604.

[0840] IR (KBr) 1744.26, 1659.82, 1599.86, 1507.20, 1450.94, 1420.55,1412.06, 1265.82, 1230.04, 1205.12, 1128.92 cm⁻¹.

Example 62N-3,4,5-Trimethoxyphenyl-3-[3-ethoxy-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0841]

[0842] To a solution of example 44 (29 mg, 0.05 mmol) in THF (3 ml)under N₂ was added a solution of 21% sodium ethoxide in ethanol (40 ul,0.1 mmol, 2 eq.). It was stirred at room temperature for 30 min, thenpartitioned between ethyl acetate and H₂O. The organic layer was washedwith brine, dried (MgSO₄), filtered and concentrated. The crude materialwas purified by flash chromatography using 1-30% acetone indichloromethane to give the title compound (4.5 mg, 16%).

[0843]¹H NMR (400 MHz, CDCl₃) δ 7.61 (t, 1H), 7.48 (d, 1H), 7.34 (s,1H), 7.31-7.20 (m, 3H), 6.87 (s, 2H), 6.59 (dd, 1H), 4.76 (q, 2H), 3.87(s, 6H), 3.78 (s, 5H), 1.56 (t, 3H) ppm.

[0844] MS(FAB) (m/z) 564.1545.

Example 63N-3,4,5-Trimethoxyphenyl-3-[3-(1-methylethoxy)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0845]

[0846] To a solution of example 44 (50 mg, 0.09 mmol) in THF (5 ml)under N₂ was added 0.95M sodium isopropoxide (100 ul, 1.2 eq.). Thereaction was stirred at room temperature for 6 h, then set in a freezerfor 17 h. Additional 0.95M sodium isopropoxide (150 ul, 1.8 eq.) wasadded, stirred for 2 h, and then partitioned between ethyl acetate andH₂O. The organic layer was washed with brine, dried (MgSO₄), filteredand concentrated. The crude material was purified by flashchromatography using 1-10% acetone in dichloromethane to give the tidecompound (9 mg, 17%).

[0847] MS(FAB) (m/z) 578.1689.

[0848] IR (KBr) 1680.23, 1623.90, 1596.74, 1508.46, 1131.81 cm⁻¹.

Example 64N-3,4,5-Trimethoxyphenyl-3-[3-(1-methoxy)isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl]phenylacetamide

[0849]

[0850] To a solution of example 44 (7 mg, 0.01 mmol) in MeOH (2.5 ml)under N₂ was added 0.95M sodium methoxide (18 ul, 2 eq.). The solutionwas stirred at room temperature for 6 h, and partitioned between ethylacetate and H₂O. The organic layer was washed with brine, dried (MgSO₄),filtered and concentrated. The crude material was purified by flashchromatography using 1-10% acetone in dichloromethane to give the tidecompound (2.5 mg, 35%).

[0851]¹H NMR (400 MHz, CDCl₃) δ 7.61 (t, 1H), 7.48 (d, 1H), 7.36 (s,1H), 7.32-7.20 (m, 4H), 6.89 (s, 2H), 6.60 (dd, 1H), 4.37 (s, 3H), 3.84(s, 6H), 3.79 (s, 3H), 3.77 (s, 2H) ppm.

[0852] MS(FAB) (m/z) 550.1387.

Example 65N-3,4,5-Trimethoxyphenyl-3-[oxazolo[4,5-c]-1,2-dihydrochloroquinolin-2-on-1-yl]phenylacetamide

[0853]

[0854] Compound from preparation 57 (122 mg, 0.226 mmol) and 2N NaOH inMeOH (2.5 ml, 5.0 mmol) were allowed to react in DMF (5 ml) at roomtemperature for 16 h. Dilution with EtOAc, extraction with H₂O (3×) andbrine, drying (MgSO₄), filtration, concentration, and purification bycolumn chromatography (silica gel, hexanes/EtOAc gradient) gave thetitle compound (74 mg, 63%).

[0855]¹H NMR (400 MHz, d₆-DMSO) d 10.36 (s, 1H), 8.95 (s, 1H), 7.55-7.45(m, 2H), 7.43-7.38 (m, 2H), 7.26 (s, 1H), 7.21 (d, 1H, J=7.3 Hz), 6.97(s, 2H), 6.57 (d, 1H, J=8.3 Hz), 3.70 (s, 2H), 3.65 (s, 6H), 3.53 (s,3H) ppm.

[0856] Mass spectrum (ES) (m/z) 520.2 [M+1].

[0857] The compounds of the invention are inhibitors of MRP1. Thus, thecompounds of the invention may be used to inhibit any neoplasm havingintrinsic and/or acquired resistance, conferred in part or in total byMRP1, to an oncolytic or oncolytics. In other words, treatment of such aneoplasm with an effective amount of a compound of this invention willcause the neoplasm to be more sensitive to chemotherapy that wasrendered less efficacious by MRP1.

[0858] Vincristine, epirubicin, daunorubicin, doxorubicin, and etoposideare oncolytics that are substrates of MRP1. See Cole, et. al.,“Pharmacological Characterization of Multidrug Resistant MRP-transfectedHuman Tumor Cells”, Cancer Research, 54:5902-5910, 1994. Since MRP1 isubiquitous in mammals, particularly humans, Nooter, K, et. al.,“Expression of the Multidrug Resistance-Associated Protein (IMP) Gene inHuman Cancers”, Clin Can. Res., 1:1301-1310, (1995), chemotherapy whosegoal is to inhibit a neoplasm employing any of those agents has thepotential to be rendered less efficacious by MRP1. Thus, neoplasms ofthe bladder, bone, breast, lung (small-cell), testis, and thyroid andmore specific types of cancer such as acute lymphoblastic andmyeloblastic leukemia, Wilm's tumor, neuroblastoma, soft tissue sarcoma,Hodgkin's and nonHodgkin's lymphomas, and bronchogenic carcinoma may beinhibited with a combination of one or more of the above oncolytics anda compound of this invention.

[0859] The biological activity of the compounds of the present inventionwas evaluated employing an initial screening assay which rapidly andaccurately measured the activity of the tested compound in inhibitingMRP1 or MDR1. Assays useful for evaluating this reversing capability arewell known in the art. See, e.g., T. McGrath, et al., BiochemicalPharmacology, 38:3611, 1989; D. Marquardt and M.S. Center, CancerResearch, 52:3157, 1992; D. Marquardt, et al., Cancer Research, 50:1426,1990; and Cole, et. al., Cancer Research, 54: 5902-5910, 1994.

[0860] Assay for Reversal of MRP1-Mediated Doxorubicin Resistance andMDR1-Mediated Vincristine Resistance: HL60/Adr and HL60/Vinc arecontinuous cell lines, which were selected for doxorubicin andvincristine resistance respectively by culturing HL60, a human acutemyeloblastic leukemia cell line, in increasing concentrations ofdoxorubicin or vincristine until a highly resistant variant wasattained.

[0861] HL60/Adr and HL60/Vinc cells were grown in RPMI 1640 (Gibco)containing 10% fetal bovine serum (FBS) and 50 μg/ml GENTAMICIN™(Sigma). Cells were harvested; washed twice with assay medium (same asculture media); counted; and diluted to 1×10⁵ cells/ml in assay medium.One hundred microliters of cells were aliquoted into wells of a 96 welltissue culture plate. Two columns of each 96 well plate served as anegative control and received assay medium containing no cells.

[0862] Test compounds and reference compounds were dissolved in dimethylsulfoxide (DMSO) at a concentration of 5 mM. Samples were diluted inassay medium and 25 μl of each test compound was added to 8 wells. Assaystandards were run in quadruplicate. Assay media was added to half ofthe wells and doxorubicin to the other half of the wells to achieve afinal volume of 150 μl per well.

[0863] The plates were incubated at 37° C. for 72 hours in a humidifiedincubator with a 5% carbon dioxide atmosphere. Cell viability andvitality was measured by oxidation of a alamarBlue™ fluorescent dyeusing standard conditions. The plates were incubated for 3 hours at 37°C. Fluorescence was determined using 550 nm excitation and 590 nmemission using a microtitre plate reader.

[0864] The ability of a test compound to reverse the resistance ofHL60/Adr and HL60/Vinc cells to doxorubicin was determined by comparisonof the absorbance of the wells containing a test compound in addition tothe oncolytic (doxorubicin) with the absorbance of wells containing theoncolytic without a test compound. Controls were used to eliminatebackground and to ensure the results were not artifactual. The resultsof the assay are expressed as percent inhibition of cell growth. Theoncolytic alone at the tested concentration minimally inhibits thegrowth of HL60/Adr or HL60/Vinc cells.

[0865] Representative compounds of formula I demonstrated a significanteffect in reversing the MRP1 multiple drug resistance. Many of thecompounds showed very significant enhancement of activity in combinationwith the oncolytic agent as opposed to the oncolytic agent alone. Inaddition, a large majority of the compounds tested displayed asignificant degree of selective inhibition of the HL60/Adr cell lineover the HL60/Vinc cell line.

[0866] When administering an oncolytic in practicing the methods of thisinvention, the amount of oncolytic employed will be variable. It shouldbe understood that the amount of the oncolytic actually administeredwill be determined by a physician, in the light of the relevantcircumstances, including the condition to be treated, the chosen routeof administration, the actual oncolytic administered, the age, weight,and response of the individual patient (mammal), and the severity of thepatient's symptoms. Of course, the amount of oncolytic administeredshould be decided and closely monitored by that patient's physician.After deciding on the oncolytic or oncolytics to employ, “ThePhysician's Desk Reference®”, published by Medical Economics Company atMontvale, N.J. 07645-1742, is a helpful resource to the physician indeciding on amounts of the oncolytic to administer and is updatedannually.

[0867] Preferred formulations, and the methods of this inventionemploying those formulations, are those which do not contain anoncolytic. Thus, it is preferred to administer the compounds of thisinvention separately from the oncolytic. The oncolytics mentioned inthis specification are commercially available and may be purchased inpreformulated forms suitable for the methods of this invention.

[0868] The compounds of formula I alone, or optionally in combinationwith an oncolytic, are usually administered in the form ofpharmaceutical formulations. These formulations can be administered by avariety of routes including oral, rectal, transdermal, subcutaneous,intravenous, intramuscular, and intranasal. Such formulations areprepared in a manner well known in the pharmaceutical art and compriseat least one active compound of formula I.

[0869] The present invention also includes methods employingpharmaceutical formulations which contain, as the active ingredient, thecompounds of formula I, and optionally an oncolytic, associated withpharmaceutical carriers. In making the formulations of the presentinvention the active ingredient(s) is usually mixed with an excipient,diluted by an excipient, or enclosed within such a carrier which can bein the form of a capsule, sachet, paper or other container. When theexcipient serves as a diluent, it can be a solid, semi-solid, or liquidmaterial, which acts as a vehicle, carrier or medium for the activeingredient. Thus, the formulations can be in the form of tablets, pills,powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions,solutions, syrups, aerosols (as a solid or in a liquid medium),ointments containing for example up to 10% by weight of the activecompound, soft and hard gelatin capsules, suppositories, sterileinjectable solutions, and sterile packaged powders.

[0870] In preparing a formulation, it may be necessary to mill theactive compound(s) to provide the appropriate particle size prior tocombining with the other ingredients. If the active compound(s) issubstantially insoluble, it ordinarily is milled to a particle size ofless than 200 mesh. If the active compound(s) is substantially watersoluble, the particle size is normally adjusted by milling to provide asubstantially uniform distribution in the formulation, e.g., about 40mesh.

[0871] Some examples of suitable excipients include lactose, dextrose,sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate,alginates, tragacanth, gelatin, calcium silicate, microcrystallinecellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulations can additionally include: lubricating agentssuch as talc, magnesium stearate, and mineral oil; wetting agents;emulsifying and suspending agents; preserving agents such as methyl- andpropylhydroxybenzoates; sweetening agents; and flavoring agents. Theformulations of the invention can be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.

[0872] The formulations are preferably formulated in a unit dosage form,each dosage containing from about 5 to about 100 mg, more usually about10 to about 30 mg, of each active ingredient. The term “unit dosageform” refers to physically discrete units suitable as unitary dosagesfor human subjects and other mammals, each unit containing apredetermined quantity of active material calculated to produce thedesired therapeutic effect, in association with a suitablepharmaceutical excipient.

[0873] The compounds of formula I are effective over a wide dosagerange. For example, dosages per day normally fall within the range ofabout 0.5 to about 30 mg/kg of body weight. In the treatment of adulthumans, the range of about 1 to about 15 mg/kg/day, in single or divideddose, is especially preferred. However, it will be understood that theamount of the compound actually administered will be determined by aphysician, in the light of the relevant circumstances, including thecondition to be treated, the chosen route of administration, the actualcompound administered, the age, weight, and response of the individualpatient, and the severity of the patient's symptoms, and therefore theabove dosage ranges are not intended to limit the scope of the inventionin any way. In some instances dosage levels below the lower limit of theaforesaid range may be more than adequate, while in other cases stilllarger doses may be employed without causing any harmful side effect,provided that such larger doses are first divided into several smallerdoses for administration throughout the day.

[0874] For preparing solid formulations such as tablets the principalactive ingredient(s) is mixed with a pharmaceutical excipient to form asolid preformulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient(s) is dispersed evenly throughout the formulation so that theformulation may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid preformulation isthen subdivided into unit dosage forms of the type described abovecontaining from 0.1 to about 500 mg of the active ingredient of thepresent invention.

[0875] The tablets or pills of the present invention may be coated orotherwise compounded to provide a dosage form affording the advantage ofprolonged action. For example, the tablet or pill can comprise an innerdosage and an outer dosage component, the latter being in the form of anenvelope over the former. The two components can be separated by entericlayer which serves to resist disintegration in the stomach and permitthe inner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids andmixtures of polymeric acids with such materials as shellac, cetylalcohol, and cellulose acetate.

[0876] The novel formulations which are liquid forms may be incorporatedfor administration orally or by injection and include aqueous solutions,suitably flavored syrups, aqueous or oil suspensions, and flavoredemulsions with edible oils such as cottonseed oil, sesame oil, coconutoil, or peanut oil, as well as elixirs and similar pharmaceuticalvehicles.

[0877] Formulations for inhalation or insufflation include solutions andsuspensions in pharmaceutical, aqueous or organic solvents, or mixturesthereof, and powders. The liquid or solid formulations may containsuitable pharmaceutical excipients as described supra. Preferably theformulations are administered by the oral or nasal respiratory route forlocal or systemic effect. Compositions in preferably pharmaceuticalsolvents may be nebulized by use of inert gases. Nebulized solutions maybe breathed directly from the nebulizing device or the nebulizing devicemay be attached to a face mask, tent, or intermittent positive pressurebreathing machine. Solution, suspension, or powder formulations may beadministered, preferably orally or nasally, from devices which deliverthe formulation in an appropriate manner.

[0878] The following formulation examples are illustrative only and arenot intended to limit the scope of the invention in any way. “Activeingredient(s)” means a compound according to formula I or apharmaceutical salt or solvate thereof optionally with one or moreoncolytics.

Formulation Example 1

[0879] Hard gelatin capsules containing the following ingredients areprepared: Quantity Ingredient (mg/capsule)N-3,4,5-trimethoxyphenyl-3-[3-azidomethyl- 30.0isoxazolo[4,5-c]-1,2-dihydro-6-chloro-quinolin-2-on-1-yl]phenylacetamide Starch 305.0 Magnesium stearate 5.0

[0880] The above ingredients are mixed and filled into hard gelatincapsules in 340 mg quantities.

Formulation Example 2

[0881] A tablet formula is prepared using the ingredients below:Quantity Ingredient (mg/capsule) N-3,4,5-trimethoxyphenyl-3-[3-methyl-25.0 isoxazolo[4,5-c]-1,2-dihydro-6-fluoro-quinolin-2-on-1-yl]phenylacetamide Cellulose, microcrystalline 200.0Colloidal silicon dioxide 10.0 Stearic acid 5.0

[0882] The components are blended and compressed to form tablets, eachweighing 240 mg.

Formulation Example 3

[0883] A dry powder inhaler formulation is prepared containing thefollowing components: Ingredient Weight %N-3,4,5-trimethoxyphenyl-3-[3-methyl- 5isoxazolo[4,5-c]-1,2-dihydro-6-methoxy-quinolin-2-on-1-yl]phenylacetamide Lactose 95

[0884] The active ingredient is mixed with the lactose and the mixtureis added to a dry powder inhaling appliance.

Formulation Example 4

[0885] Tablets, each containing 30 mg of active ingredient, are preparedas follows: Quantity Ingredient (mg/capsule)N-3,4,5-trimethoxyphenyl-3-[3-methyl- 30.0 mgisoxazolo[4,5-c]-1,2-dihydro-5- (4-methoxyphenyl)-quinolin-2-on-1-yl]phenylacetamide Starch 45.0 mg Microcrystalline cellulose 35.0 mgPolyvinylpyrrolidone 4.0 mg (as 10% solution in water) Sodiumcarboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc 1.0 mg Total120 mg

[0886] The active ingredient, starch and cellulose are passed through aNo. 20 mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders, which are thenpassed through a 16 mesh U.S. sieve. The granules so produced are driedat 50-60° C. and passed through a 16 mesh U.S. sieve. The sodiumcarboxymethyl starch, magnesium stearate, and talc, previously passedthrough a No. 30 mesh U.S. sieve, are then added to the granules which,after mixing, are compressed on a tablet machine to yield tablets eachweighing 120 mg.

Formulation Example 5

[0887] Capsules, each containing 40 mg of medicament are made asfollows: Quantity Ingredient (mg/capsule)N-3,4,5-trimethoxyphenyl-3-[3-methyl- 40.0 mgisoxazolo[4,5-c]-1,2-dihydro-6-carboxy-quinolin-2-on-1-yl]phenylacetamide Starch 109.0 mg Magnesium stearate1.0 mg Total 150.0 mg

[0888] The active ingredient, cellulose, starch, and magnesium stearateare blended, passed through a No. 20 mesh U.S. sieve, and filled intohard gelatin capsules in 150 mg quantities.

Formulation Example 6

[0889] Suppositories, each containing 25 mg of active ingredient aremade as follows: Ingredient AmountN-3,4,5-trimethoxypbenyl-3-[3-((2-methoxy- 25 mg1-benzyl-ethan-2-al)amidyl)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin- 2-on-1-yl]phenylacetamideSaturated fatty acid glycerides to 2,000 mg

[0890] The active ingredient is passed through a No. 60 mesh U.S. sieveand suspended in the saturated fatty acid glycerides previously meltedusing the minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2.0 g capacity and allowed to cool.

Formulation Example 7

[0891] Suspensions, each containing 50 mg of medicament per 5.0 ml doseare made as follows: Ingredient AmountN-3,4,5-trimethoxyphenyl-3-[3-phenylthio- 50.0 mgisoxazolo[4,5-c]-1,2-dihydro-6- chloroquinolin-2-on-1-yl]phenylacetamideXanthan gum 4.0 mg Sodium carboxymethyl cellulose (11%) 50.0 mgMicrocrystalline cellulose (89%) Sucrose 1.75 g Sodium benzoate 10.0 mgFlavor and Color q.v. Purified water to 5.0 ml

[0892] The active ingredient, sucrose and xanthan gum are blended,passed through a No. 10 mesh U.S. sieve, and then mixed with apreviously made solution of the microcrystalline cellulose and sodiumcarboxymethyl cellulose in water. The sodium benzoate, flavor, and colorare diluted with some of the water and added with stirring. Sufficientwater is then added to produce the required volume.

Formulation Example 8

[0893] Capsules, each containing 15 mg of medicament, are made asfollows: Quantity Ingredient (mg/capsule)N-3,4,5-trimethoxyphenyl-3-[3-methyl- 15.0 mgisoxazolo[4,5-c]-1,2-dihydro-5- (4-trifluoromethylphenyl)-quinolin-2-on-1-yl]phenylacetamide Starch 407.0 mg Magnesium stearate 3.0 mg Total425.0 mg

[0894] The active ingredient, cellulose, starch, and magnesium stearateare blended, passed through a No. 20 mesh U.S. sieve, and filled intohard gelatin capsules in 425 mg quantities.

Formulation Example 9

[0895] An intravenous formulation may be prepared as follows: IngredientQuantity N-3,4,5-trimethoxyphenyl-3-[3-methyl- 250.0 mgisoxazolo[4,5-c]-1,2-dihydro-6- iodoquinolin-2-on-1-yl]phenylacetamideIsotonic saline 1000 ml

Formulation Example 10

[0896] A topical formulation may be prepared as follows: IngredientQuantity N-3,4,5-trimethoxyphenyl-3-[3-methyl- 1-10 gisoxazolo[4,5-c]-1,2-dihydro-6-(thien-2-yl)-quinolin-2-on-1-yl]phenylacetamide Emulsifying Wax 30 g Liquid Paraffin20 g White Soft Paraffin to 100 g

[0897] The white soft paraffin is heated until molten. The liquidparaffin and emulsifying wax are incorporated and stirred untildissolved. The active ingredient is added and stirring is continueduntil dispersed. The mixture is then cooled until solid.

Formulation Example 11

[0898] Sublingual or buccal tablets, each containing 10 mg of activeingredient, may be prepared as follows: Quantity Ingredient (mg/capsule)N-3,4,5-trimethoxyphenyl-3-[3- 10.0 mg (1-methoxy)-isoxazolo[4,5-c]-1,2-dihydro-6-chloroquinolin-2-on-1-yl] phenylacetamide Glycerol 210.5 mg Water 143.0 mg  Sodium Citrate  4.5 mg Polyvinyl Alcohol 26.5 mgPolyvinylpyrrolidone 15.5 mg Total 410.0 mg 

[0899] The glycerol, water, sodium citrate, polyvinyl alcohol, andpolyvinylpyrrolidone are admixed together by continuous stirring andmaintaining the temperature at about 90° C. When the polymers have goneinto solution, the solution is cooled to about 50-55° C. and the activeingredient is slowly admixed. The homogenous mixture is poured intoforms made of an inert material to produce a drug-containing diffusionmatrix having a thickness of about 24 mm. This diffusion matrix is thencut to form individual tablets having the appropriate size.

[0900] Another preferred formulation employed in the methods of thepresent invention employs transdermal delivery devices (“patches”). Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of the compounds of the present invention in controlledamounts. The construction and use of transdermal patches for thedelivery of pharmaceutical agents is well known in the art See, e.g.,U.S. Pat. No. 5,023,252, issued Jun. 11, 1991, herein incorporated byreference. Such patches may be constructed for continuous, pulsatile, oron demand delivery of pharmaceutical agents.

[0901] Frequently, it will be desirable or necessary to introduce thepharmaceutical formulation to the brain, either directly or indirectly.Direct techniques usually involve placement of a drug delivery catheterinto the host's ventricular system to bypass the blood-brain barrier.One such implantable delivery system, used for the transport ofbiological factors to specific anatomical regions of the body, isdescribed in U.S. Pat. No. 5,011,472, issued Apr. 30, 1991, which isherein incorporated by reference.

[0902] Indirect techniques, which are generally preferred, usuallyinvolve formulating the compositions to provide for drug latentiation bythe conversion of hydrophilic drugs into lipid-soluble drugs orprodrugs. Latentiation is generally achieved through blocking of thehydroxy, carbonyl, sulfate, and primary amine groups present on the drugto render the drug more lipid soluble and amenable to transportationacross the blood-brain barrier. Alternatively, the delivery ofhydrophilic drugs may be enhanced by intra-arterial infusion ofhypertonic solutions, which can transiently open the blood-brainbarrier.

We claim:
 1. A compound of formula I:

where: het is a five (5) membered heteroaryl ring containing N and asecond heteroatom selected from N, O, or S; wherein the non-fused carbonatom of the heteroaryl ring is optionally substituted with C₁-C₆ alkyl,aryl, substituted aryl, heterocycle, substituted heterocycle, an aminoacid ester, CH₂OH, CH₂O-heterocycle, halo, CH₂N₃, CH₂SR¹, CH₂NR⁴R⁵, OR¹,SR¹², S(CH₂)_(n)-phenyl, or NR⁴R⁵; provided that when het is pyrazole orimidazole, the saturated nitrogen of the het ring is optionallysubstituted with C₁-C₄ alkyl; R is (CH₂)_(m′)CHR¹NHR², O(CH₂)₂NHR²,(CH₂)_(m′)COR³, NHR², and (CH₂)_(m′)CHR¹NR⁴R⁵; R′ is hydrogen, hydroxy,or O(C₁-C₆ alkyl optionally substituted with phenyl or C₃-C₇cycloalkyl); m and m′ are independently at each occurrence 0, 1, or 2;R¹ is independently at each occurrence hydrogen or C₁-C₆ alkyl; R² ishydrogen, COR⁶, CH₂R⁶, SO₂R⁷, or a moiety of the formula

R³ is hydrogen, hydroxy, C₁-C₆ alkoxy, an amino acid ester, an aminoacid, or NR⁴R⁵; R⁴ is hydrogen or C₁-C₆ alkyl; R⁵ is hydrogen, C₁-C₆alkyl, C₆-C₁₀ bicycloalkyl, (C₁-C₄ alkyl)-phenyl, (C₁-C₄ alkyl)-CO₂R₁,CH₂CO₂R¹, aryl, substituted aryl, (CH₂)_(n)CHR⁸NHC(O)OC(CH₃)₃,(CH₂)_(n)NH₂, (CH₂)₂NHCOR⁶, (CH₂)₂OR¹, (CH₂)_(q)-heterocycle,(CH₂)_(q)-substituted heterocycle, or R⁴ and R⁵, together with thenitrogen to which they are attached, combine to form a pyrrolidin-1-yl,piperidin-1-yl, hexamethyleneimin-1-yl, or morpholin-4-yl ring; n is 1,2, 3, or 4; q is 0, 1, 2, or 3; R⁶ is C₁-C₆ alkyl, substituted C₃-C₆cycloalkyl, aryl, substituted aryl, tert-butoxy, (CH₂)_(q)-heterocycle,(CH₂)_(q)-substituted heterocycle, (CH₂)_(n)S(O)_(r)R¹, C(CH₃)₂CH₂N(R¹),(CH₂)_(n)CHR⁸NHC(O)OC(CH₃)₃, (CH₂)_(n)CHR⁸NH₂, (CH₂)₂NH-aryl, or NHR⁷;R^(6′) is C₁-C₆ alkyl, substituted C₃-C₆ cycloalkyl, aryl, substitutedaryl, (CH₂)_(q)-heterocycle, (CH₂)_(q)-substituted heterocycle,(CH₂)_(n)S(O)_(r)R¹, C(CH₃)₂CH₂N(R¹)₂, (CH₂)_(n)CHR⁸NH—C(O)OC(CH₃)₃,(CH₂), CHR⁸NH₂, or (CH₂)₂NH-aryl; r is 0, 1, or 2; R⁷ is C₁-C₆ alkyl,phenyl, or substituted phenyl; R⁸ is hydrogen or CO₂R¹; and R⁹, R¹⁰, andR¹¹ are independently at each occurrence hydrogen, halo, CO₂R¹, aryl,substituted aryl, thiophene, C₁-C₄ alkoxy, (C₁-C₃ alkyl)phenyl, or C₂-C₆alkenyl; R¹² is C₁-C₆ alkyl, (C₁-C₄ alkyl)-phenyl, aryl, substitutedaryl, heterocycle or substituted heterocycle; or a pharmaceutical saltthereof; provided that if R⁹ and R¹⁰ are hydrogen and R¹¹ is chloro,then het is not


2. The compound according to claim 1 where m is 0 and R is at the metaposition.
 3. The compound according to claim 2 where R is(CH₂)_(m′)CHR¹NHR² and m′ is 0 and R¹ is methyl.
 4. The compoundaccording to claim 3 where R² is 3,4,5-trimethoxybenzyl.
 5. The compoundaccording to claim 2 where R is (CH₂)_(m′)COR³ and m′ is 0 or
 1. 6. Thecompound according to claim 5 where R³ is (3,4,5-trimethoxyphenyl)amino,(4-aminosulfonylphenyl)amino, or (6-methoxyquinolin-8-yl)amino.
 7. Thecompound according to claim 2 where R is (CH₂)_(m′)CHR¹NR⁴R⁵ and m′ is0, and R¹ and R⁴ is hydrogen.
 8. The compound according to claim 7 whereR⁵ is 5-methylisoxazol-3-yl, 3,5-dimethoxy-4-hydroxybenzyl, or3,4,5-trimethoxybenzyl.
 9. A method of inhibiting MRP1 in a mammal whichcomprises administering to a mammal in need thereof an effective amountof a compound of formula I, as defined in claim 1, or a pharmaceuticalsalt thereof.
 10. The method according to claim 9 where the mammal is ahuman.
 11. The method according to claim 10 where the compound offormula I is a compound where m is 0 and R is at the meta position. 12.The method according to claim 11 where the compound of formula I is acompound where R is (CH₂)_(m′)CHR¹NHR² and m′ is 0 and R¹ is methyl. 13.The method according to claim 12 where the compound of formula I is acompound where R² is 3,4,5-trimethoxybenzyl.
 14. The method according toclaim 11 where the compound of formula I is a compound where R is(CH₂)_(m′)COR³ and m′ is 0 or
 1. 15. The method according to claim 14where the compound of formula I is a compound where R³ is(3,4,5-trimethoxyphenyl)amino, (4-aminosulfonylphenyl)amino, or(6-methoxyquinolin-8-yl)amino.
 16. The method according to claim 11where the compound of formula I is a compound where R is(CH₂)_(m′)CHR¹NR⁴R⁵ and m′ is 0, and R¹ and R⁴ is hydrogen.
 17. Themethod according to claim 16 where the compound of formula I is acompound where R⁵ is 5-methylisoxazol-3-yl,3,5-dimethoxy-4-hydroxybenzyl, or 3,4,5-trimethoxybenzyl.
 18. A methodof inhibiting a resistant neoplasm, or a neoplasm susceptible toresistance, in a mammal which comprises administering to a mammal inneed thereof an effective amount of a compound of formula I, as definedin claim 1, or a pharmaceutical salt thereof; in combination with aneffective amount of one or more oncolytic agents.
 19. The methodaccording to claim 18 where the mammal is a human.
 20. The methodaccording to claim 19 where the oncolytic(s) is selected from:doxorubicin, daunorubicin, epirubicin, vincristine, and etoposide. 21.The method according to claim 19 where the neoplasm is of the Wilm'stype, bladder, bone, breast, lung (small-cell), testis, or thyroid orthe neoplasm is associated with acute lymphoblastic and myeloblasticleukemia, neuroblastoma, soft tissue sarcoma, Hodgkin's andnon-Hodgkin's lymphomas, and bronchogenic carcinoma.
 22. The methodaccording to claim 19 where the compound of formula I is a compoundwhere m is 0 and R is at the meta position.
 23. The method according toclaim 22 where the compound of formula I is a compound where R isCHR¹NHR² and R¹ is methyl.
 24. The method according to claim 23 wherethe compound of formula I is a compound where R² is3,4,5-trimethoxybenzyl.
 25. The method according to claim 22 where thecompound of formula I is a compound where R is COR³ or (CH₂)COR³. 26.The method according to claim 25 where the compound of formula I is acompound where R³ is (3,4,5-trimethoxyphenyl)amino,(4-aminosulfonylphenyl)amino, or (6-methoxyquinolin-8-yl)amino.
 27. Themethod according to claim 22 where the compound of formula I is acompound where R is (CH₂)NR⁴R⁵ and R⁴ is hydrogen.
 28. The methodaccording to claim 27 where the compound of formula I is a compoundwhere R⁵ is 5-methylisoxazol-3-oyl, 3,5-dimethoxy-4-hydroxybenzyl, or3,4,5-trimethoxybenzyl.
 29. A pharmaceutical formulation comprising acompound of formula I, as defined in claim 1, or a pharmaceutical saltthereof; in combination with one or more pharmaceutical carriers,diluents, or excipients therefor.
 30. A pharmaceutical formulationcomprising: (a) a compound of formula I, as defined in claim 1, or apharmaceutical salt thereof; (b) one or more oncolytic agents; and (c)one or more pharmaceutical carriers, diluents, or excipients therefor.31. The formulation according to claim 30 where the oncolytic(s) isselected from: doxorubicin, daunorubicin, epirubicin, vincristine, andetoposide.
 32. A use of a compound of formula I, as defined in claim 1,or a pharmaceutically acceptable salt thereof, for the manufacture of amedicament for inhibiting a resistant neoplasm, or a neoplasmsusceptible to resistance in a mammal.
 33. A use of a compound offormula I, as defined in claim 1, or a pharmaceutically acceptable saltthereof, for the manufacture of a medicament for inhibiting MRP1.
 34. Ause of a compound of formula I, as defined in claim 1, or apharmaceutically acceptable salt thereof, for the manufacture of amedicament for inhibiting MRP1 conferred MDR in a resistant neoplasm, ora neoplasm susceptible to resistance in a mammal.
 35. A use of acompound of formula I, as defined in claim 1, in therapy.
 36. Apharmaceutical composition for inhibiting MRP1 in a mammal whichcomprises an effective amount of a compound of formula I, as defined inclaim 1, or a pharmaceutical salt thereof.
 37. The composition accordingto claim 36 where the mammal is a human.
 38. The composition accordingto claim 37 where the compound of formula I is a compound where m is 0and R is at the meta position.
 39. The composition according to claim 38where the compound of formula I is a compound where R is(CH₂)_(m′)CHR¹NHR² and m′ is 0 and R¹ is methyl.
 40. The compositionaccording to claim 39 where the compound of formula I is a compoundwhere R² is 3,4,5-trimethoxybenzyl.
 41. The composition according toclaim 38 where the compound of formula I is a compound where R is(CH₂)_(m′)COR³ and m′ is 0 or
 1. 42. The composition according to claim41 where the compound of formula I is a compound where R³ is(3,4,5-trimethoxyphenyl)amino, (4-aminosulfonylphenyl)amino, or(6-methoxyquinolin-8-yl)amino.
 43. The composition according to claim 38where the compound of formula I is a compound where R is(CH₂)_(m′)CHR¹NR⁴R⁵ and m′ is 0, and R¹ and R⁴ is hydrogen.
 44. Thecomposition according to claim 43 where the compound of formula I is acompound where R⁵ is 5-methylisoxazol-3-yl,3,5-dimethoxy-4-hydroxybenzyl, or 3,4,5-trimethoxybenzyl.
 45. Apharmaceutical composition for inhibiting a resistant neoplasm, or aneoplasm susceptible to resistance, in a mammal which comprisesadministering to a mammal in need thereof an effective amount of acompound of formula I, as defined in claim 1, or a pharmaceutical saltthereof; in combination with an effective amount of one or moreoncolytic agents.
 46. The composition according to claim 45 where themammal is a human.
 47. The composition according to claim 46 where theoncolytic(s) is selected from: doxorubicin, daunorubicin, epirubicin,vincristine, and etoposide.
 48. The composition according to claim 46where the neoplasm is of the Wilm's type, bladder, bone, breast, lung(small-cell), testis, or thyroid or the neoplasm is associated withacute lymphoblastic and myeloblastic leukemia, neuroblastoma, softtissue sarcoma, Hodgkin's and non-Hodgkin's lymphomas, and bronchogeniccarcinoma.
 49. The composition according to claim 46 where the compoundof formula I is a compound where m is 0 and R is at the meta position.50. The composition according to claim 49 where the compound of formulaI is a compound where R is CHR¹NHR² and R¹ is methyl.
 51. Thecomposition according to claim 50 where the compound of formula I is acompound where R² is 3,4,5-trimethoxybenzyl.
 52. The compositionaccording to claim 49 where the compound of formula I is a compoundwhere R is COR³ or (CH₂)COR³.
 53. The composition according to claim 52where the compound of formula I is a compound where R³ is(3,4,5-trimethoxyphenyl)amino, (4-aminosulfonylphenyl)amino, or(6-methoxyquinolin-8-yl)amino.
 54. The composition according to claim 49where the compound of formula I is a compound where R is (CH₂)NR⁴R⁵ andR⁴ is hydrogen.
 55. The composition according to claim 54 where thecompound of formula I is a compound where R⁵ is 5-methylisoxazol-3-oyl,3,5-dimethoxy-4-hydroxybenzyl, or 3,4,5-trimethoxybenzyl.